Team:Peking S/lab/notebook/lwh
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Wenhan Luo's Notebook
summary
In our project I cloned genes of A-factor synthase afsA and A-factor receptor arpA,and I also standardized these two genes .Besides, I also constructed and characterized quorum sensing genes named cinI and cinR. I further characterized its feasibilities in constructing synthetic microbial consortia.
Contents
6.24-7.5 Learn the basic experiment skills including transformation and ligation and digestion.
7.6-7.7 Ask professor Honglong some questions about Streptomyces and acquire the bacteria strain from him.
7.8-7.12 Try to PCR an a-factor generator gene(afsa) and its corresponding receivers, the repessor gene called arpa from Streptomyces.
7.12-7.16 Using different enzymes(prime star、esay pfu、TAG ) in PCR to amplification afsa gene and arpa gene from the Streptomyces .
7.17-7.23 No matter which DNA polymerase we used,we can not obtain those two genes, so we try to change the anneal temperature in PCR process.
7.24-7.28 I got afsa and arpa eventually and going to construct a-factor chemical-wire gene circuit.
7.29-8.18 Constuct a three input logic gate whose input is CAI-I and Arabinose and Salicylate and output is green fluorescence protein.
8.19-9.19 Do a ribosome binding site mutation of an AND GATE whose input is AHL and Salicylate.
9.20-10.1 Make a characterization of the salicylate(sal), induce a gene circuit which consists of the sal promoter and a gfp gene by different concentration and different time span.Get the fluorescence intensity cell by cell using FCM(flow cytometry) and depict the graph.