畜牧与饲料科学2016，Vol.37Issue(2)：29-33,5.
牛肌酸激酶同功酶CK-BB的全基因合成及其原核表达
Whole Gene Synthesis and Prokaryotic Expression of Bovine Creatime Kinase Isoenzyme CK-BB
摘要
Abstract
The creatine kinase plays an important role in bovine energy metabolism. When inflammation occurs, there will be a locally-specific increase of CK. According, the creatine kinase may serve as a potential marker of bovine diseases. In this study, a total of 54 mutually overlapped primers were designed and synthesized according to the published amino acid sequence of bovine creatime kinase isoenzyme CK-BB and the codon usage preferred by E. coli (BL21). A total of 6 DNA fragments of CK-BB gene, including CK-BB1, CK-BB2, CK-BB3, CK-BB4, CK-BB5 and CK-BB6, were synthesized by assembled PCR assay. Using the mixture of the 6 fragments as template, the second round PCR was conducted with head primer ck-1 and tail primer ck-54, and the amplifying product was connected with vector pET28b. The positive recombinants were subsequently identified, cloned and sequenced. A correct synthetic CK-BB gene was obtained by proofreading PCR assay, which lays a foundation for large-scale preparation of bovine isoenzyme CK-BB.关键词
牛肌酸激酶同功酶/密码子优化/全基因合成Key words
bovine creatime kinase isoenzyme/codon optimization/whole gene synthesis分类
农业科技引用本文复制引用
杨鼎,吴江鸿,祁云霞,赵世华..牛肌酸激酶同功酶CK-BB的全基因合成及其原核表达[J].畜牧与饲料科学,2016,37(2):29-33,5.基金项目
内蒙古农牧业科学院青年创新基金（2013QN JJM11）。 （）
