Allopatric integrations selectively change host transcriptomes, leading to varied expression efficiencies of exotic genes in Myxococcus xanthus

Microbial Cell Factories

Research

Yue-Zhong Li¹  Li-Ping Zhu¹  Kui Han¹  Zhi-Feng Li¹  Hai-Long Wang¹  You-Ming Zhang¹  Xiu-Nan Yi¹  Xin-Jing Yue¹  Lian-Shuai Zheng¹ 

[1] State Key Laboratory of Microbial Technology, School of Life Science, Shandong University, 250100, Jinan, China;

关键词: Transposition;    Site-directed integration;    Chloramphenicol acetyl transferase;    Epothilone biosynthetic gene cluster;    Expression efficiency;    Transcriptome;    Myxococcus xanthus;   

DOI  :  10.1186/s12934-015-0294-5

 received in 2015-05-17, accepted in 2015-07-07,  发布年份 2015

来源: Springer

【 摘 要 】

BackgroundExotic genes, especially clustered multiple-genes for a complex pathway, are normally integrated into chromosome for heterologous expression. The influences of insertion sites on heterologous expression and allotropic expressions of exotic genes on host remain mostly unclear.ResultsWe compared the integration and expression efficiencies of single and multiple exotic genes that were inserted into Myxococcus xanthus genome by transposition and attB-site-directed recombination. While the site-directed integration had a rather stable chloramphenicol acetyl transferase (CAT) activity, the transposition produced varied CAT enzyme activities. We attempted to integrate the 56-kb gene cluster for the biosynthesis of antitumor polyketides epothilones into M. xanthus genome by site-direction but failed, which was determined to be due to the insertion size limitation at the attB site. The transposition technique produced many recombinants with varied production capabilities of epothilones, which, however, were not paralleled to the transcriptional characteristics of the local sites where the genes were integrated. Comparative transcriptomics analysis demonstrated that the allopatric integrations caused selective changes of host transcriptomes, leading to varied expressions of epothilone genes in different mutants.ConclusionsWith the increase of insertion fragment size, transposition is a more practicable integration method for the expression of exotic genes. Allopatric integrations selectively change host transcriptomes, which lead to varied expression efficiencies of exotic genes.

【 授权许可】

CC BY   
© Zhu et al. 2015