|
|
| Line 161: |
Line 161: |
| | </p> | | </p> |
| | | | |
| − |
| |
| − |
| |
| − | <html>
| |
| − | <script>
| |
| − | ////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////
| |
| − |
| |
| − | $(document).ready(function () {
| |
| − |
| |
| − | $("#HQ_page").attr('id', '');
| |
| − |
| |
| − |
| |
| − | //highlight current page on the menu
| |
| − | highlight_current_page_menu();
| |
| − |
| |
| − | //accessing submenus
| |
| − | $(".menu_item").click(function () {
| |
| − | $(".submenu_control_icon", this).toggleClass("open");
| |
| − | $(this).next(".submenu").fadeToggle(400);
| |
| − | });
| |
| − |
| |
| − | //mobile menu access
| |
| − | $(".igem_2018_team_mobile_bar").click(function () {
| |
| − | $(this).next().toggleClass("displaying_menu");
| |
| − | });
| |
| − |
| |
| − | });
| |
| − |
| |
| − |
| |
| − | function highlight_current_page_menu() {
| |
| − |
| |
| − | var page_url = "https://2018.igem.org/";
| |
| − | page_url = page_url + wgPageName;
| |
| − | $("a[href$='" + page_url + "']").children().addClass("current_page");
| |
| − |
| |
| − | //if the page is in a submenu, open the submenu and make the appropiate changes
| |
| − | if ($(".current_page").hasClass("submenu_item")) {
| |
| − |
| |
| − | $(".current_page").parent().parent().fadeToggle(400);
| |
| − | $(".current_page").parent().parent().prev().addClass("current_page");
| |
| − | $(".menu_item.current_page > .submenu_control_icon").toggleClass("open");
| |
| − |
| |
| − | }
| |
| − | }
| |
| − |
| |
| − |
| |
| − | ////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////
| |
| − | </script>
| |
| − |
| |
| − |
| |
| − | <style>
| |
| − | /**************************************************************************************************************************************************************************************************/
| |
| − |
| |
| − |
| |
| − |
| |
| − | /**************************************************************************************************************************************************************************************************/
| |
| − | /* DEFAULT WIKI SETTINGS */
| |
| − | /**************************************************************************************************************************************************************************************************/
| |
| − |
| |
| − |
| |
| − | #home_logo, #sideMenu { display:none; }
| |
| − | #sideMenu, #top_title, .patrollink {display:none;}
| |
| − | #content { margin-left:0px; margin-top:-7px; padding:0px; width:100%;}
| |
| − | body {background-color:white; }
| |
| − | #bodyContent h1, #bodyContent h2, #bodyContent h3, #bodyContent h4, #bodyContent h5 { margin-bottom: 0px; }
| |
| − |
| |
| − | .judges-will-not-evaluate { border: 4px solid #e4dede; padding: 2% !important; width: 92%!important;}
| |
| − |
| |
| − |
| |
| − | /**************************************************************************************************************************************************************************************************/
| |
| − | /* MENU */
| |
| − | /**************************************************************************************************************************************************************************************************/
| |
| − |
| |
| − | /*this wraps the whole of the menu*/
| |
| − | .igem_2018_team_menu {
| |
| − | background-color:#c4baba;
| |
| − | border-left: 1px solid #c4baba;
| |
| − | display:block;
| |
| − | float:right;
| |
| − | height:100vh;
| |
| − | max-width: 270px;
| |
| − | overflow-y: auto;
| |
| − | overflow-x: hidden;
| |
| − | padding:0px;
| |
| − | position:fixed;
| |
| − | right:0%;
| |
| − | text-align:left;
| |
| − | width: 15%;
| |
| − | }
| |
| − |
| |
| − |
| |
| − | .igem_2018_team_menu.displaying_menu{
| |
| − | display:block;
| |
| − | }
| |
| − |
| |
| − | .igem_2018_team_menu a {
| |
| − | color: #484848;
| |
| − | text-decoration:none;
| |
| − | }
| |
| − |
| |
| − | .igem_2018_team_menu img {
| |
| − | width:100%;
| |
| − | }
| |
| − |
| |
| − |
| |
| − | .igem_2018_team_menu .menu_item {
| |
| − | background-color: #c4baba;
| |
| − | border-bottom: 1px solid #928b8b;
| |
| − | clear: both;
| |
| − | color: #484848;
| |
| − | cursor: pointer;
| |
| − | float: left;
| |
| − | font-size: 120%;
| |
| − | font-weight: bold;
| |
| − | padding: 15px 0px 15px 5%;
| |
| − | width: 100%;
| |
| − | }
| |
| − |
| |
| − | .igem_2018_team_menu .menu_item.direct_link {
| |
| − | color: #484848;
| |
| − | padding-left: 15%;
| |
| − | }
| |
| − | .igem_2018_team_menu .menu_item:hover {
| |
| − | background-color: #ecb656;
| |
| − | }
| |
| − |
| |
| − |
| |
| − | .igem_2018_team_menu .menu_item .submenu_control_icon {
| |
| − | color: #484848;
| |
| − | float: left;
| |
| − | width: 10%;
| |
| − | }
| |
| − |
| |
| − | /* submenu icon "+" "-"*/
| |
| − | .igem_2018_team_menu .menu_item .submenu_control_icon::before {
| |
| − | content: "+";
| |
| − | }
| |
| − |
| |
| − | /* submenu icon "+" "-"*/
| |
| − | .igem_2018_team_menu .menu_item .submenu_control_icon.open::before {
| |
| − | content: "-";
| |
| − | }
| |
| − |
| |
| − | /*submenu wrapper*/
| |
| − | .igem_2018_team_menu .submenu{
| |
| − | background-color: #e4dede;
| |
| − | clear:both;
| |
| − | display:none;
| |
| − | float: left;
| |
| − | width:100%;
| |
| − | }
| |
| − |
| |
| − | /*styling for a submenu item*/
| |
| − | .igem_2018_team_menu .submenu .submenu_item {
| |
| − | border-bottom: 1px solid #c4baba;
| |
| − | color: #635d5d;
| |
| − | height: 30px;
| |
| − | float: left;
| |
| − | font-size: 110%;
| |
| − | font-weight: bold;
| |
| − | padding: 12px 0px 0px 15%;
| |
| − | width: 100%;
| |
| − | }
| |
| − |
| |
| − | .igem_2018_team_menu .submenu .submenu_item:hover {
| |
| − | background-color: #f3bd5d;
| |
| − | }
| |
| − |
| |
| − |
| |
| − | .igem_2018_team_menu .submenu .submenu_item.current_page,
| |
| − | .igem_2018_team_menu .menu_item.current_page,
| |
| − | .igem_2018_team_menu .menu_item.direct_link.current_page {
| |
| − | background-color:#7acbd8;
| |
| − | }
| |
| − |
| |
| − |
| |
| − |
| |
| − | /*mobile menu bar styling*/
| |
| − | /**************************************************************************************************************************************************************************************************/
| |
| − |
| |
| − |
| |
| − | .igem_2018_team_mobile_bar {
| |
| − | background-color:#e4dede;
| |
| − | border-bottom: 1px solid #c4baba;
| |
| − | cursor:pointer;
| |
| − | display:none;
| |
| − | float:left;
| |
| − | margin-top: 0;
| |
| − | padding: 5px 0;
| |
| − | position:fixed;
| |
| − | width:100%;
| |
| − | }
| |
| − |
| |
| − | .igem_logo_mobile img {
| |
| − | width:70px;
| |
| − | }
| |
| − |
| |
| − | .igem_logo_mobile {
| |
| − | float:left;
| |
| − | padding-left: 5%;
| |
| − | width: 30%;
| |
| − | }
| |
| − |
| |
| − | .igem_menu_control_mobile img {
| |
| − | width:25px;
| |
| − | }
| |
| − |
| |
| − | .igem_menu_control_mobile {
| |
| − | float:right;
| |
| − | padding-right:5%;
| |
| − | padding-top:5px;
| |
| − | text-align:right;
| |
| − | width: 30%;
| |
| − | }
| |
| − |
| |
| − |
| |
| − |
| |
| − | /**************************************************************************************************************************************************************************************************/
| |
| − | /* CONTENT OF THE PAGE */
| |
| − | /**************************************************************************************************************************************************************************************************/
| |
| − |
| |
| − | /* general wrapper for the content */
| |
| − | .igem_2018_team_content {
| |
| − | background-color:white;
| |
| − | display:block;
| |
| − | width: 87%;
| |
| − | }
| |
| − |
| |
| − | /* subwrapper to center content */
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper {
| |
| − | margin:auto;
| |
| − | max-width: 1400px;
| |
| − | width:90%;
| |
| − | }
| |
| − |
| |
| − |
| |
| − |
| |
| − | /*general styling*/
| |
| − | /**************************************************************************************************************************************************************************************************/
| |
| − |
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper h1 { font-size: 210%;}
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper h2 { font-size: 190%;}
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper h3 { font-size: 170%;}
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper h4 { font-size: 150%;}
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper h5 { font-size: 140%;}
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper h6 { font-size: 130%;}
| |
| − |
| |
| − |
| |
| − | /* styling for the titles h1, h2*/
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper h1, .igem_2018_team_content .igem_2018_team_column_wrapper h2 {
| |
| − | border-bottom:0px;
| |
| − | color: #635d5d;
| |
| − | font-family: "Arial Black", Gadget, sans-serif;
| |
| − | padding: 10px 0px;
| |
| − | }
| |
| − |
| |
| − | /* styling for the titles h3, h3, h5, h6 */
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper h3,
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper h4,
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper h5,
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper h6 {
| |
| − | border-bottom:0px;
| |
| − | color: #928b8b;
| |
| − | font-family: "Arial Black", Gadget, sans-serif;
| |
| − | padding: 5px 0px;
| |
| − | }
| |
| − |
| |
| − | /* text */
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper p {
| |
| − | font-size: 130%;
| |
| − | font-family: Arial, Helvetica, sans-serif;
| |
| − | padding: 5px 0px;
| |
| − | text-align: left;
| |
| − | color: #484848;
| |
| − | }
| |
| − |
| |
| − | /* Links */
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper a {
| |
| − | color: #5bc7d8;
| |
| − | font-weight: bold;
| |
| − | text-decoration: underline;
| |
| − | text-decoration-color:#5bc7d8;
| |
| − | transition: all 0.4s ease;
| |
| − | -webkit-transition: all 0.4s ease;
| |
| − | -moz-transition: all 0.4s ease;
| |
| − | -ms-transition: all 0.4s ease;
| |
| − | -o-transition: all 0.4s ease;
| |
| − | }
| |
| − |
| |
| − | /* hover for the links */
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper a:hover {
| |
| − | color: #f8b732;
| |
| − | text-decoration:none;
| |
| − | }
| |
| − |
| |
| − |
| |
| − | /* Table */
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper table {
| |
| − | border: 1px solid #928b8b;
| |
| − | border-collapse: collapse;
| |
| − | font-size: 130%;
| |
| − | width: 100%;
| |
| − | }
| |
| − |
| |
| − | /* table cells */
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper td {
| |
| − | border: 1px solid #c4baba;
| |
| − | border-collapse: collapse;
| |
| − | font-size: 105%;
| |
| − | padding: 10px;
| |
| − | vertical-align: text-top;
| |
| − | }
| |
| − |
| |
| − | /* table headers */
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper th {
| |
| − | background-color:#c4baba;
| |
| − | border: 1px solid #928b8b;
| |
| − | border-collapse: collapse;
| |
| − | font-size: 110%;
| |
| − | padding: 10px;
| |
| − | vertical-align: text-top;
| |
| − | }
| |
| − |
| |
| − |
| |
| − |
| |
| − | /* non numbered lists */
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper ul, .igem_2018_team_content .igem_2018_team_column_wrapper ol {
| |
| − | font-size: 130%;
| |
| − | font-family: Arial, Helvetica, sans-serif;
| |
| − | padding:0px 20px;
| |
| − | }
| |
| − |
| |
| − |
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper ul ul li, .igem_2018_team_content .igem_2018_team_column_wrapper ul ul ul li,
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper ul ol li, .igem_2018_team_content .igem_2018_team_column_wrapper ul ul ol li,
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper ol ol li, .igem_2018_team_content .igem_2018_team_column_wrapper ul ol ul li,
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper ol ul li, .igem_2018_team_content .igem_2018_team_column_wrapper ul ol ol li,
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper ol ul ul li, .igem_2018_team_content .igem_2018_team_column_wrapper ol ol ul li,
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper ol ol ol li, .igem_2018_team_content .igem_2018_team_column_wrapper ol ul ol li{ font-size: 76%; }
| |
| − |
| |
| − |
| |
| − |
| |
| − | /*layout classes*/
| |
| − | /**************************************************************************************************************************************************************************************************/
| |
| − |
| |
| − | /*main layout class */
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper .column {
| |
| − | float:left;
| |
| − | margin: 1% 2%;
| |
| − | padding: 0px;
| |
| − | }
| |
| − |
| |
| − | /* 100% */
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper .column.full_size { width:96%; }
| |
| − |
| |
| − | /* 66% */
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper .column.two_thirds_size { width: 62.6%; }
| |
| − |
| |
| − | /* 33% */
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper .column.third_size { width: 29.3%; }
| |
| − |
| |
| − |
| |
| − |
| |
| − |
| |
| − | /*styling for all images*/
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper .column.full_size img,
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper .column.two_thirds_size img,
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper .column.third_size img {
| |
| − | margin-bottom: 15px;
| |
| − | width: 100%;
| |
| − | }
| |
| − |
| |
| − |
| |
| − | /* page break */
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper .clear {
| |
| − | clear:both;
| |
| − | }
| |
| − | /*add extra space to page break with clear class*/
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper .clear.extra_space {
| |
| − | height: 30px;
| |
| − | }
| |
| − |
| |
| − | /* horizontal line to divide the page*/
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper .line_divider {
| |
| − | border-top: 1px solid #c4baba;
| |
| − | margin: auto;
| |
| − | width: 98%;
| |
| − | }
| |
| − |
| |
| − |
| |
| − |
| |
| − |
| |
| − | /*support classes*/
| |
| − | /**************************************************************************************************************************************************************************************************/
| |
| − |
| |
| − |
| |
| − | /*Button */
| |
| − | /************************************************/
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper .button_link {
| |
| − | font-size: 130%;
| |
| − | margin: 30px auto;
| |
| − | text-align: center;
| |
| − | }
| |
| − |
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper .button_link a {
| |
| − | background-color: #5bc7d8;
| |
| − | color: #635d5d !important;
| |
| − | font-weight: bold;
| |
| − | margin: auto;
| |
| − | text-decoration: none !important;
| |
| − | padding: 10px 15px;
| |
| − | }
| |
| − |
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper .button_link a:hover {
| |
| − | background-color: #f8b732 !important;
| |
| − | }
| |
| − |
| |
| − |
| |
| − |
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper .highlight {
| |
| − | padding: 15px 20px;
| |
| − | }
| |
| − |
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper .highlight p,
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper .highlight h1,
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper .highlight h2,
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper .highlight h3,
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper .highlight h4,
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper .highlight h5,
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper .highlight h6 {
| |
| − | padding: 5px 15px;
| |
| − | }
| |
| − |
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper .highlight.decoration_background {
| |
| − | background-color: #e4dede;
| |
| − | }
| |
| − |
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper .highlight.decoration_A_top {
| |
| − | border-top: 4px solid #5bc7d8;
| |
| − | }
| |
| − |
| |
| − |
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper .highlight.decoration_A_full {
| |
| − | border: 4px solid #5bc7d8;
| |
| − | }
| |
| − |
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper .highlight.decoration_B_top {
| |
| − | border-top: 4px solid #f8b732
| |
| − | }
| |
| − |
| |
| − |
| |
| − | .igem_2018_team_content .igem_2018_team_column_wrapper .highlight.decoration_B_full {
| |
| − | border: 4px solid #f8b732;
| |
| − | }
| |
| − |
| |
| − |
| |
| − |
| |
| − |
| |
| − | /*mobile*/
| |
| − | /**************************************************************************************************************************************************************************************************/
| |
| − |
| |
| − |
| |
| − | /* 1800px */
| |
| − | /************************************************/
| |
| − | @media only screen and (max-width: 1800px) {
| |
| − | .igem_2018_team_content { width: 85%;}
| |
| − | .igem_2018_team_menu {display:block;}
| |
| − | }
| |
| − |
| |
| − | /* 1400px */
| |
| − | /************************************************/
| |
| − | @media only screen and (max-width: 1400px) {
| |
| − | .igem_2018_team_menu .menu_item { font-size:100%;}
| |
| − | .igem_2018_team_menu .submenu .submenu_item { font-size:90%;}
| |
| − | .igem_2018_team_menu {display:block;}
| |
| − | }
| |
| − |
| |
| − | @media only screen and (max-width: 1001px) {
| |
| − | .igem_2018_team_menu {display:block;}
| |
| − | }
| |
| − |
| |
| − | /* 1000px */
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| − | /************************************************/
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In the following article, we describe the process of building our model. During the construction, we aim to build a model by combining the data fitting principle in mathematical modeling with the help of MATLAB through multiple experiments, and obtain a relatively successful fitting formula after multiple optimization, so as to reflect the data trend reasonably.
During the experiment, we first controlled the consistency of the concentration of the bacterial liquid so that it would not affect the experimental results. There are 3 groups of experimental statistic, they record 3 kinds of mutant statistic——The independent variable—concentration of lactic acid(lactate)，time of reaction(t), the dependent variable—intensity of fluorescence(Fluorescence). On the basis of reasonable hypothesis and correlation coefficient test, according to the fitting of known data, we get the function and function curve that can reflect data trend reasonably. This curve illustrates the functional relationship between the fluorescence intensity of the dependent variable and the independent variable.
1: According to some reasonable assumptions given in professional knowledge, we believe that the linear function may be a non-linear function between the dependent variable and the independent variable, since the addition of lactic acid and IPTG will tend to influence the growth rate of the organism, and the lactic acid into the cell and the ai-2 generated by the Luxs catalysis also need to be of a certain and unequal time, so that the whole reaction process does not constitute a linear function requirement. So it's a logarithmic function or a polynomial function.
2: Under each reasonable hypothesis, MATLAB is used to perform corresponding fitting. According to the correlation coefficient R to test the fitting effect, the fitting function formula that can reasonably reflect the data trend can be obtained after continuous optimization.
t=0 min Relative Fluorescence= 1.496 t 3 -8.148 t 2 + 10.31 t + 8.919 R2= 0.1473
t=5 min Relative Fluorescence= 9.304 t 3 -49.68 t 2 + 63.56 t + 22.25 R2= 0.9464
t=10 min Relative Fluorescence= 1.286 t 3 -12.36 t 2 + 19.78 t + 29.03 R2= 0.9986
t=20 min Relative Fluorescence= 17.26 t 3 -83.37 t 2 + 96.5 t + 19.91 R2= 0.9957
t=30 min Relative Fluorescence= 9.669 t 3 - 48.15 t 2 + 58.23 t + 39.34 R2= 0.8645
t=60 min Relative Fluorescence= 20.96 t 3 - 106.2 t 2 + 134.9 t + 42.16 R2= 0.9482
t=5 min Relative Fluorescence= 6.669 t 3 -38.13 t 2 + 55.27 t + 13.89 R2= 0.9808
t=10 min Relative Fluorescence= -3.422 t 3 +7.808 t 2 + 8.072 t + 11.18 R2=0.9946
t=30 min Relative Fluorescence= 7.02 t 3 - 37.92 t 2 + 55.03 t + 9.921 R2= 0.9998
t=60 min Relative Fluorescence= 9.403 t 3 - 55.39 t 2 + 80.41 t + 8.451 R2= 0.9787
t=5 min Relative Fluorescence= -2.332 t 3 +8.402 t 2 + 2.239 t + 20.74 R2= 0.9834
t=10 min Relative Fluorescence= 0.4742 t 3 -1.957 t 2 + 5.059 t +26.94 R2= 0.9544
t=30 min Relative Fluorescence= -7.451 t 3 +36.49 t 2 -44.01 t + 39.25 R2= 0.9645
t=60 min Relative Fluorescence= -0.7971 t 3 +4.694 t 2 -13.22 t + 39.88 R2= 0.9761
t=10 min Relative Fluorescence= 12.03 t^3 -62.83 * t^2 +86.66 * t+10.36 R2= 0.7222
t=30 min Relative Fluorescence= 6.297 t 3 -35.63 t 2 + 53.2 t + 9.274 R2= 0.8867
t=60 min Relative Fluorescence= 9.122 * t 3 - 51.26 * t 2 + 73.33 * t + 5.924 R2= 0.9866
t=5 min Relative Fluorescence= -12.48 t 3 +45.65 t 2 -26.91 t + 18.86 R2= 0.9762
t=10 min Relative Fluorescence= -1.135 t 3 +5.337 t 2 -3.825 t + 23.12 R2= 0.511
t=30 min Relative Fluorescence= 3.63 t 3 - 25.57 t 2 + 40.87 t + 14.54 R2= 0.893
t=60 min Relative Fluorescence= 9.562 t 3 - 53.63 t 2 + 74.38 t + 9.515 R2= 0.9907
When [IPTG]=1 mM , The concentration of fluorescence at each point of time shows a similar trend towards the change in the rate of lactate, which is increased by the increase in lactate concentration, and then the fluorescence starts to go up and then down, and then there's a tendency to increase the intensity of the fluorescence, at the end of the level of the lactic acid, which is at the peak of the lactic acid, which is at the level of 0.5 to the one, and the fluorescence is at a peak. According to actual production, the concentration of yogurt in fermentation cannot exceed 1mM[1]. Therefore, the engineered bacteria can monitor the lactic acid concentration of yogurt during fermentation. In terms of time, when IPTG was added with 1mm, the fluorescence value of 5-60 min was all higher than that of 0 min, indicating that the reaction of engineering bacteria to lactic acid was effective during this period. Therefore, 5 min was selected as the appropriate reaction time. Because the fluorescence intensity of fiber detection can be stable within 200 milliseconds, the actual production can be obtained, and the time for testing the lactic acid concentration by using this engineering bacteria is 5 min.
When [IPTG]=1 mM, with the increase of time, the intensity of fluorescence expressed by engineered bacteria all lower than the intensity at 0 min. This may be due to the fact that at 0 min, the expression of lldR has not yet started or the expression quantity is too low, so the opening of lldPRD operon promoter caused by the introduction of engineering bacteria is not prevented. However, as the reaction time increases, lldR gradually produces, and the GFP background expression decreases due to the opening of lldPRD operon promoter. From the perspective of time, when the reaction time was 5 min, the change trend of fluorescence intensity with the concentration of lactic acid was consistent with the initial reaction time, so the good reaction time of engineering bacteria could be achieved within 5 min.
When there is no lactose operon, different concentrations of lactic acid are added. Although the engineered bacteria will still react according to different concentrations of lactic acid, the curve of the function is obviously irregular with different reaction times.
lldPRD operon promoter-GFP & T7- lldPRD operon promoter-GFP& Lldr- T7-lldPRD operon promoter-GFP:
We fitted the change of fluorescence intensity expression of the three engineered bacteria at the reaction time of 5 min, hoping to obtain the difference of the three engineered bacteria.
In the process of production of yogurt, the lactic acid content should not exceed 1 mM. So when the lactic acid content was lower than 1 mM, the lactose operon or lldR is added to the engineered bacteria and the lactic acid concentration is lower than 1 mM, indicating that the two engineered bacteria can achieve high sensitivity. Besides, when lactic acid content was lower than 1 mM, the lactose operon or lldR is added to the engineered bacteria is higher than those which only have lldPRD operon, indicating that both the lactose operon and lldR have an improved effect on the initial engineered bacteria.(BBa_K822000 ). Compared with the improvement of BBa_K822000 by lldR, it is obvious that the addition of lactose operon makes engineering bacteria more sensitive.
Except for the beginning of the reaction, The rest of the functional curves showed similar trends, especially when the reaction time was 5 min and 10 min, and the peak value was reached before the concentration of lactic acid was 1 mM. It proved that the engineering bacteria was best used to detect the lactic acid concentration in yogurt at the reaction time of 5 min and 10 min. In order to reduce the reaction time of engineered bacteria, the reaction time of 5 min was selected.
According to the result of fitting, after 30 min of reaction, the engineered bacteria were highly sensitive to different concentrations of lactic acid. However, the reacting time was to long, the increase of fluorescence value may be caused by the death of bacteria, which has no reference value.
lldPRD operon promoter-Luxs-Lldr × LsrA promoter-GFP & lldPRD operon promoter-Luxs × LsrA promoter-GFP& Lldr- T7-lldPRD operon promoter-GFP
We fitted the change of fluorescence intensity expression of the three engineered bacteria at the reaction time of 5 min, hoping to obtain the difference of the three engineering bacteria.
Apparently, when the lactic acid concentration is less than 1 mM, included lldPRD operon promoter - Luxs - Lldr x LsrA promoter - the GFP and Lldr - T7 has - lldPRD operon promoter - GFP, two kinds of engineering bacteria in the peak, and the sensitivity of the former than the latter. This indicates that when the lactic acid concentration of yogurt is not over the limit, these two types of engineered bacteria can produce higher sensitivity, and the addition of the swarm sensing system can improve the sensitivity of engineered bacteria to the lactic acid concentration. The engineering bacteria contains lldPRD operon promoter - Luxs x LsrA promoter - GFP in lactic acid concentration is lower than 2.5 mM, to produce higher sensitivity.
All in all, from what has been discussed above, we can learn that contains lldPRD operon promoter - Luxs - Lldr x LsrA promoter - GFP engineering bacteria is applied to we have the best equipment engineering bacteria.
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