Difference between revisions of "Team:JNFLS/Design"
| Line 5: | Line 5: | ||
<h1>Design</h1> | <h1>Design</h1> | ||
<p> | <p> | ||
| − | In the very beginning of our project, we brainstormed about the detection measures for HCV. The current detection method used in all blood centers in Shandong province is ELISA-based HCV antibody detection, which caused the awkward situation that not even a single positive result was gained in the last decade. HCV antibody detection will be the past, but what the future will be? ELISA-based antigen detection? No, the detection must be sufficiently sensitive, what if the amount of virus in the blood is too low? Nucleic acid detection? No, it shares the same drawback as the previous method; also, RNA is way easier to be degraded. In the end, the works of Zhang et al and Zhou inspired us. We eventually designed a biosensor based on aptamer and rolling circle amplification. | + | In the very beginning of our project, we brainstormed about the detection measures for HCV. The current detection method used in all blood centers in Shandong province is ELISA-based HCV antibody detection, which caused the awkward situation that not even a single positive result was gained in the last decade. HCV antibody detection will be the past, but what the future will be? ELISA-based antigen detection? No, the detection must be sufficiently sensitive, what if the amount of virus in the blood is too low? Nucleic acid detection? No, it shares the same drawback as the previous method; also, RNA is way easier to be degraded. In the end, the works of Zhang et al.[3] and Zhou[4] inspired us. We eventually designed a biosensor based on aptamer and rolling circle amplification. |
</p> | </p> | ||
<img src="https://static.igem.org/mediawiki/2018/b/b3/T--JNFLS--ban.jpg"style="width:70%"> | <img src="https://static.igem.org/mediawiki/2018/b/b3/T--JNFLS--ban.jpg"style="width:70%"> | ||
| − | <p>After the experiment began, we were in urge to find an efficient way to gain HCV core protein for SELEX of aptamers. HCV core protein originally consists of 191 amino acid, which is hard to be expressed in prokaryotic cells. However, Li [1] and Wu [2] found in their studies that HCV core protein can be expressed more rapidly and operably if the protein is truncated into 119, 125, or 173 amino acid in size. The HCVC protein we used consists of 120AA and 173AA, based on the works mentioned above. The result showed that the amount of protein expressed increase. | + | <p>After the experiment began, we were in an urge to find an efficient way to gain HCV core protein for SELEX of aptamers. HCV core protein originally consists of 191 amino acid, which is hard to be expressed in prokaryotic cells. However, Li [1] and Wu [2] found in their studies that HCV core protein can be expressed more rapidly and operably if the protein is truncated into 119, 125, or 173 amino acid in size. The HCVC protein we used consists of 120AA and 173AA, based on the works mentioned above. The result showed that the amount of protein expressed increase. |
</p> | </p> | ||
<img src="https://static.igem.org/mediawiki/2018/d/d7/T--JNFLS--tian.jpg"style="width:80%"> | <img src="https://static.igem.org/mediawiki/2018/d/d7/T--JNFLS--tian.jpg"style="width:80%"> | ||
Revision as of 13:33, 9 October 2018
Design
In the very beginning of our project, we brainstormed about the detection measures for HCV. The current detection method used in all blood centers in Shandong province is ELISA-based HCV antibody detection, which caused the awkward situation that not even a single positive result was gained in the last decade. HCV antibody detection will be the past, but what the future will be? ELISA-based antigen detection? No, the detection must be sufficiently sensitive, what if the amount of virus in the blood is too low? Nucleic acid detection? No, it shares the same drawback as the previous method; also, RNA is way easier to be degraded. In the end, the works of Zhang et al.[3] and Zhou[4] inspired us. We eventually designed a biosensor based on aptamer and rolling circle amplification.
After the experiment began, we were in an urge to find an efficient way to gain HCV core protein for SELEX of aptamers. HCV core protein originally consists of 191 amino acid, which is hard to be expressed in prokaryotic cells. However, Li [1] and Wu [2] found in their studies that HCV core protein can be expressed more rapidly and operably if the protein is truncated into 119, 125, or 173 amino acid in size. The HCVC protein we used consists of 120AA and 173AA, based on the works mentioned above. The result showed that the amount of protein expressed increase.
- Li Shengtao. Purification of expression of HCV "truncated" core protein (HCV Core125) and preparation of antibodies [D]. Kunming university of technology,2012.
- Wu Xianbo. Cloning, expression and purification and antigenicity analysis of HCV core antigen gene fragments [D]. First military medical university of the people's liberation army,2003.
- Zhang Songbai, Zheng Liying, Hu xia, Shen Guangyu, Liu Xuewen, Shen Guoli, Yu Ruqin. High sensitivity detection of thrombin by fluorescent adaptor sensor based on competitive trigger rolling ring amplification [J]. Analytical chemistry,2015,43(11):1688-1694.
- Zhou hui. Study on nucleic acid aptamer sensor based on signal amplification triggered by competitive mechanism [D]. Hunan university,2009.
