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| 微小牛蜱Bm91基因的克隆及其真核表达载体的构建 | |
| 引用本文： | 王玙,曾巧英,罗建勋,马米玲,关贵全,殷宏. 微小牛蜱Bm91基因的克隆及其真核表达载体的构建[J]. 甘肃农业大学学报, 2009, 44(1) |
| 作者姓名： | 王玙 曾巧英 罗建勋 马米玲 关贵全 殷宏 |
| 作者单位： | 1. 甘肃农业大学动物医学院,甘肃,兰州,730070;中国农业科学院兰州兽医研究所,家畜疫病病原生物学国家重点实验室,甘肃省动物寄生虫病重点实验室,甘肃,兰州,730046 2. 甘肃农业大学动物医学院,甘肃,兰州,730070 3. 中国农业科学院兰州兽医研究所,家畜疫病病原生物学国家重点实验室,甘肃省动物寄生虫病重点实验室,甘肃,兰州,730046 |
| 基金项目： | 国家高技术研究发展计划(863计划)，国家自然资源平台项目，国家支撑计划 |
| 摘 要： | 参照GenBank中收录的微小牛蜱Bm91基因的核苷酸序列,设计了1对特异性引物,从我国微小牛蜱半饱血雌性成蜱肠道和唾液腺研磨物中提取总RNA,利用M-MLV RTase cDNA Synthesis Kit合成双链cDNA,并采用PCR技术扩增获得的Bm91基因,将其克隆到pGEM-T Easy载体中并进行PCR、酶切和测序分析鉴定.结果表明:所克隆的Bm91基因序列与GenBank上收录的Bm91基因序列的同源性达97.1%,而且该片段含有完整的开放阅读框;将该基因从克隆载体上用EcoRⅠ单酶切消化并亚克隆到巴斯德毕赤酵母分泌型表达载体pP1C9K的EcoRⅠ酶切位点,再次进行PCR、酶切和测序分析鉴定,结果表明成功构建并获得了微小牛蜱Bm91基因的重组真核表达载体pP1C9K-Bm91. |
| 关 键 词： | 微小牛蜱 Bm91基因 真核表达载体 |
Cloning of Bm91 gene of Boophilus microplus and construction of eukaryotic expression vector with Bm91 gene |
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| WANG Yu,ZENG Qiao-ying,LUO Jian-xun,MA Mi-ling,GUAN Gui-quan,YIN Hong. Cloning of Bm91 gene of Boophilus microplus and construction of eukaryotic expression vector with Bm91 gene[J]. Journal of Gansu Agricultural University, 2009, 44(1) | |
| Authors: | WANG Yu ZENG Qiao-ying LUO Jian-xun MA Mi-ling GUAN Gui-quan YIN Hong |
| Affiliation: | WANG Yu1,2,ZENG Qiao-ying1,LUO Jian-xun2,MA Mi-ling2,GUAN Gui-quan2,YIN Hong2(1.College of Veterinary Medicine,Gansu Agricultural University,Lanzhou 730070,China,2.Key Laboratory of Veterinary Parasitology of Gansu Province,State Key Laboratory of Veterinary Etiological Biology,Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China) |
| Abstract: | The total RNA was extracted from the salivary glands and the intestine of semiengorged adult female Boophilus microplus and double cDNA was synthesized using M-MLV RTase cDNA Synthesis Kit.According to the nucleotide sequences of Bm91 published in GenBank,a pair of primers was designed and a 1 908 bp Bm91 gene was amplified by PCR.The target gene was subcloned into pGEM-T Easy vector and tested by PCR,enzyme digestion and sequencing.The sequencing showed that the cloned Bm91 gene shared 97.1 %homology with ... |
| Keywords: | Boophilus microplus Bm91 gene recombinant eukaryotic expression vector |
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