Team:Peking S/lab/notebook/fjc

Yanrong Ma's Notebook

summary

Cloned genes of A-factor synthase afsA and its 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

Molecular cloning training . such as transformation and DNA ligation and Enzyme digestion.

7.6-7.7

Obtaining Streptomyces strain from professor Honglong and acquired some skills in Streptomyces culturing.

7.8-7.12

Usinig PCR strategy to get a-factor generator gene afsA and a-factor receptor gene arpa from Streptomyces genome.

7.13-7.23

Using different polymerases(prime star、esay pfu、TAG ) in PCR to gain afsa and arpa genes from the Streptomyces strain. And we finally got those two genes by changing many PCR parameters.

7.24-8.13

Constructing the A-factor receive system including constitutively expressed arpA and a gfp whose expression is controlled by arpA and afsA synthesized molecules.

8.14-9.5

Testing A-factor system performance.

9.6-9.18

Construction cinI/cinR system mainly contains constitutively expressed cinR and a gfp controlled by cinR and cinI synthesized chemical molecules.

9.19-10.1

Characterization of cinI/cinR system.