Team:SBS SH 112144/Basic Part

Basic Parts

Promoter: BBa_J23100

Description: Parts J23100, a common and popular promoter in E.coli protein expression, present in plasmid J61002. This basic part belongs to a family of constitutive promoter, J23100 through J23119, which can be functioned to initiate the transcription of our cyanophage lysozyme DNA sequence.

Length: 35 bp

RBS: BBa_B0034

Description: This RBS is based on Elowitz(1999) repressiliator. In the previous experimentation, the efficiency of RBS BBa_B0034 is defined as 1.0.

Length: 12 bp

Coding Sequence: Cyanophage Lysozyme Gene BBa_K2888000

Description: This cyanophage lysozyme discovered by Heideburg et al in Yellowstone National Park {Heideburg:2009hb} can target to and specifically lyse the cell wall of cyanobacteria by cooperating with Bugbuster chemical solution. Among the 27 lysozyme gene family, this cyanophage lysozyme gene gives the most efficient and effective soluble expression as well as purification. Unlike other bacteria, cyanobacteria have much thicker layers which are difficult to be lysed with common enzymes. Therefore, cyanobacteria-targeted lysozyme and other chemicals breaking through these unique barriers are needed in chemical lysis process.

Length: 546 bp

Coding Sequence: mlrA Gene BBa_K2888001

Description: This mlrA gene is a part of the microcystin degradation system discovered in Sphingopyxis sp. C-1. In our experiment, we intend to deal with Microcystin-LR, the most common type of microcystin toxin produced in cyanobacteria. The mlrA gene is responsible for hydrolytic cleavage and linearization of cyclic structure of MCLR.

Length: 1014 bp

Terminator BBa_B0015

center

Description: Terminator BBa_B0015 is a double terminator consisting of BBa_B0010 and BBa_B0012.This is the most commonly used terminator. It seems to be reliable.

Length: 129 bp

6x His tag

Description:Description: We add 6xhistag into the coding region by PCR. Facilitating purification, 6xhistag severs as a marked purification tag for Ni-NTA column. Moreover, 6xhistag also helps expressed enzymes immobilize as well as bind to our designed prototype device.