摘要
为了构建奶牛溶菌酶(lysozyme,LYZ)原核表达载体,试验人工设计并合成LYZ基因CDS序列,将其克隆至表达载体p ET32T,转化大肠杆菌TOP10,筛选阳性菌株提取质粒,对重组质粒进行PCR扩增、双酶切及测序鉴定。结果表明:PCR扩增和双酶切产物的分子质量分别为1 000 bp和400 bp,与预期大小一致;扩增产物经测序鉴定,也与目的片段完全一致。说明原核表达载体LYZp ET32T构建成功。
The aim of the present study was to construct prokaryotic expression vector of Bos taurus lysozyme(LYZ) gene.The CDS sequence of LYZ gene was designed and synthesized,then cloned into the expression vector p ET32 T and transformed to E.coli TOP10.The positive clones were screened to extract plasmid,and the recombinant plasmid was identified by PCR,restriction analysis and sequencing.The results showed that the molecular sizes of PCR amplification and double enzyme digestion were 1 000 bp and 400 bp,respectively.These were consistent with the expected molecular size.The amplification products were sequenced and identical with the target fragments.The results indicated that the Bos taurus lysozyme prokaryotic expression vector(LYZ-p ET32 T) was successfully constructed.
作者
李云龙
高峰
张海涛
顾开朗
LI Yunlong, GAO Feng, ZHANG Haitao, GU Kailang(Xuzhou Vocational College of Bioengineering, Xuzhou 221006, Chin)
出处
《黑龙江畜牧兽医》
CAS
北大核心
2018年第5期11-13,261,共4页
Heilongjiang Animal Science And veterinary Medicine
基金
徐州市科技计划项目(KC15N0012)
关键词
奶牛溶菌酶
质粒
大肠杆菌
克隆
奶牛
原核表达载体
Bos taurus lysozyme
plasmid
Escherichia coli
cloning
dairy cattle
prokaryotic expression vector