摘要
为了克隆微小牛蜱Bm86 基因及构建该基因的表达载体,以微小牛蜱饥饿幼蜱的破解物提取的总RNA为模板,参照已发表的微小牛蜱Bm86基因的核苷酸序列,设计了1对引物,采用RT-PCR技术获得微小牛蜱Bm86基因;将Bm86基因克隆入载体,并进行序列分析,结果证明,克隆的Bm86基因序列与GenBank上登录的Bm86基因序列的同源性达97.4%,而且该序列包含完整的开放阅读框。将该基因克隆入真核表达载体 pPIC9K,构建并获得了重组真核表达载体pPIC9K-Bm86。
The total RNA was abstracted from the larvae of Boophilus microplus, and a 1982bp Bm86 gene was amplified by RT-PCR. The target gene was subcloned into T vector. The sequencing showed that the nucleotide sequence of the cloned Bm86 gene shared 97.4% homology with the data published in (GenBank) and this fragment contained the complete open reading frame of Bm86 gene. After digesting of the (T-Bm86) and the expression vector pPIC9K with EcoRⅠ and NotⅠ, restrictively, the Bm86 gene was inserted into the digested vector pPIC9K. The identification by PCR, digestion and sequencing indicated that the construction of the recombinant eukaryotic expression vector was successful and designated pPIC9K-Bm86.
出处
《中国兽医科技》
CSCD
北大核心
2005年第2期124-128,共5页
Chinese Journal of Veterinary Science and Technology
基金
国家高技术研究发展计划(863)项(2003AA241110)
国家自然科学基金资助项目(30270992)
关键词
微小牛蜱
Bm86基因
克隆
重组表达载体
Boophilus microplus
Bm86 gene
cloning
recombinant express vector
