摘要
根据GenBank上发表的猪附红细胞体全基因组序列(登录号:NC_015153.1),针对其中的信号识别颗粒(SRP)基因设计合成一对特异性引物,应用PCR方法扩增猪附红细胞体信号识别颗粒54(SRP54)基因片段,克隆至pMD18-T simple载体上,经PCR、酶切初步鉴定正确后进行测序,对所克隆片段进行生物信息学分析,并构建重组表达质粒pGEX-SRP54。结果显示,克隆的SRP54基因片段大小为1 164bp,与GenBank中参考序列的同源性为97%;该基因编码381个氨基酸,编码蛋白的等电点为6.51,无跨膜区;重组表达质粒pGEX-SRP54构建正确。
According to the complete genome sequence of Eperythrozoon suis published in GenBank（NC_ 015153.1）,a pair of specific primers were designed and synthetized against the signal recognition particle gene sequence. The signal recognition particle 54 gene fragment of Eperythrozoon suis was amplified by PCR and cloned to pMD18-T simple. After preliminary identification correctly by PCR and enzyme digestion,sequencing was conducted. Then bioinformatics analyseis was conducted, and the recombinant expression plasmid pGEX-SRP54 was constructed. The results showed that the gene fragment Of SRP54 was 1 164 bp, and 97% homology with corresponding sequences in GenBank; It encoded 381 amino acids and the isoelectric point was 6.51. There was no transmembrane area; The recombinant expression plasmid of pGEX-SRP54 was constructed correctly, which laid the foundation for the follow-up studies.
出处
《动物医学进展》
CSCD
北大核心
2011年第12期37-40,共4页
Progress In Veterinary Medicine
基金
国家自然科学基金项目(30860203)
公益性行业(农业)科研专项项目(200903036-13)
关键词
猪附红细胞体
信号识别颗粒
克隆
生物信息学分析
表达质粒
Eperythrozoon suis
signal recognition particle
clone
bioinformatics analysis
expressionplasmid
