摘要
根据捻转血矛线虫Hc38基因产物(登录号AY749124)保守结构域所在核酸序列设计1对引物,该引物包含Kozak序列、HindⅢ酶切位点、XbaⅠ酶切位点。以含有捻转血矛线虫Hc38基因保守结构域片段的pMD19-T为模板,经PCR扩增获得Hc38基因保守结构域片段,用HindⅢ、XbaⅠ双酶切该片段,回收后将此基因片段克隆至相同酶切回收后的真核表达载体pcDNA3.1(+)中,获得重组质粒pcD-NA3.1-Hc38。经酶切分析、序列测定和PCR鉴定,证实了重组质粒的正确性。
According to Hc38 gene(GenBank number: AY749124)of Haemonchus contortus, a pair of primers with Kozak sequence, HindⅢ and Xba Ⅰ sits were densigned. The fragment in conserved domain was amplified by PCR from pMD19T-Hc38. The PCR product was digested with HindⅢ and Xba Ⅰ ,then inserted into pcDNA3.1 (+) between Hind Ⅲ and Xba Ⅰ sites to generate an expression plasmid pcDNA3.1 Hc38. The recombinant plasmid was identified by enzyme digestion,PCR and sequence. The eukaryotic ex pression plasmid pcDNA3.1-Hc38 was successfully constructed.
出处
《动物医学进展》
CSCD
北大核心
2009年第2期22-25,共4页
Progress In Veterinary Medicine
基金
新疆兵团科技局社会发展项目(2006GG31)