文献类型：期刊文献

英文题名：Available methods for assembling expression cassettes for synthetic biology

作者：Wang, Tianwen[3];Ma, Xingyuan[1,2];Zhu, Hu[4];Li, Aitao[1,2];Du, Guocheng[3];Chen, Jian[3]

机构：[1]E China Univ Sci & Technol, Sch Biotechnol, Shanghai 214122, Peoples R China;[2]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 214122, Peoples R China;[3]Jiangnan Univ, State Key Lab Food Sci & Technol, Wuxi 214122, Jiangsu, Peoples R China;[4]China Univ Petr E China, Ctr Bioengn & Biotechnol, Qingdao 266555, Peoples R China

年份：2012

卷号：93

期号：5

起止页码：1853

外文期刊名：APPLIED MICROBIOLOGY AND BIOTECHNOLOGY

收录：;EI(收录号：20121114848350);WOS:【SCI-EXPANDED(收录号:WOS:000300657900006)】；

基金：Grants from the National Science Foundation of China (31000054, 30873190) and the Fundamental Research Funds for the Central Universities (No. JUSRP10917) supported this research.

语种：英文

外文关键词：Synthetic biology; Simultaneous expression; Pathway construction; Yeast recombination; Cre-loxP site-specific recombination; Acembl system

摘要：Studies in the structural biology of the multicomponent protein complex, metabolic engineering, and synthetic biology frequently rely on the efficient over-expression of these subunits or enzymes in the same cell. As a first step, constructing the multiple expression cassettes will be a complicated and time-consuming job if the classic and conventional digestion and ligation based cloning method is used. Some more efficient methods have been developed, including (1) the employment of a multiple compatible plasmid expression system, (2) the rare-cutter-based design of vectors, (3) in vitro recombination (sequence and ligation independent cloning, the isothermally enzymatic assembly of DNA molecules in a single reaction), and (4) in vivo recombination using recombination-efficient yeast (in vivo assembly of overlapping fragments, reiterative recombination for the chromosome integration of foreign expression cassettes). In this review, we systematically introduce these available methods.