密码子优化BTRCP-CypA融合基因的PCR合成  

The Codon Optimized BTRCP-CypA Gene Synthesized by PCR

作　　者：孟祥平[1] 白雪飞[1] 杨建英[1] 乔晓岚[1] Meng Xiangping;Bai Xuefei;Yang Jianying;Qiao Xiaolan(Medical Technology and Engineering College of Henan University of Science and Technology, Luoyang, 471000)

机构地区：[1]河南科技大学医学技术与工程学院,洛阳471000

出　　处：《基因组学与应用生物学》2017年第11期4445-4450,共6页Genomics and Applied Biology

基　　金：国家自然科学基金项目(No.31201119);河南省教育厅自然科学研究项目(No.14A310026)共同资助

摘　　要：为了提高BTRCP-Cyp A融合基因的表达量,对基因密码子进行优化,主要是去除过多富含AT碱基以及提高GC含量。在此基础上提出一种用于基因合成的PCR法。拆分单链寡核首酸长度为59 nt左右,重叠区碱基数14~21 nt,Tm值为46~62℃。具体步骤如下:把拆分的寡核首酸组装成300~500 bp左右的中间片段,再将中间片段拼接成全长基因。结果表明利用该法法合成了BTRCP-Cyp A融合基因,并提高了表达量。因而改良的PCR法能经济、简便、高效地进行基因合成,具有良好的应用前景。In order to improve the expression level ofBTRCP-CypA gene, we optimized schemes for BTRCP-CypA in accordance with bias of codon to remove unstable factors such as AT-rich sequences and improve G＋C content. On this basis, we set up a PCR method for gene synthesis. The split oligonucleotides length was about 59 nt. The base number in over lapped region was 14-21 nt. Tm was 46-62℃. The concrete steps of this method were as fol- lows： Assembling split oligonucleotides into 300-500 bp length of intermediate blocks and splicing the intermediate blocks into the full length gene. Results showed that BTRCP-CypA was synthesized and the expression level of BTRCP-CypA was greatly increased. Therefore, it is an economic, simple, rapid and effective method for gene syn- thesis, as well as has the potential application value in the field of molecular biology.

关 键 词：基因合成 PCR 化学合成 密码子优化 

分 类 号：Q78[生物学—分子生物学]