摘要
目的克隆广西菲牛蛭水蛭素基因的cDNA,并对该基因以及氨基酸进行序列分析。方法根据已发表的马尼拉菲牛蛭水蛭素基因cDNA设计一对引物,从广西菲牛蛭的头部提取总RNA后,采用RT-PCR扩增cDNA,将产物克隆至载体PMD-19T,PCR鉴定后进行测序。结果广西菲牛蛭水蛭素基因cDNA序列长度为251 bp,编码框由83个氨基酸组成,其中包括由20个氨基酸的信号肽,以及63个氨基酸组成的成熟肽。广西菲牛蛭水蛭素基因的氨基酸序列和已报道的广东菲牛蛭、马尼拉菲牛蛭水蛭素基因HM1、HM2基因的氨基酸序列相比较,同源性分别为94%,91.8%,84.7%。结论广西菲牛蛭水蛭素基因cDNA序列长度为251 bp,由83个氨基酸组成。
Purpose To clone the cDNA from Hirudinaria manillensis of Guangxi and to analyze its sequence and amino acids.Methods A pair of primers were designed based on the published Hirudinaria manillensis genes.The total RNA was extracted from the head of Hirudinaria manillensis of Guangxi.The cDNA was amplified by RT-PCR using primers and cloned into the pMD 19-T Vector.Plasmid DNAs obtained from positive clones were identified by means of PCR reaction.The sequences of cDNA were determined with positive clones directly.Results The sequence analysis showed that the cDNA sequence was 251 bp which encoded an open reading frame（ORF） of 83 amino acids,also included a signal peptide of 20 amino acids,and mature protein of 63 amino acids.The amino acids sequence had a homology with Hirudinaria manillensis of Guangdong of 94%,HM1 of 91.8%,HM2 of 84.7%.Conclusion The cDNA sequence of Hirudinaria manillensis from Guangxing was 251 bp and included 83 amino acids.
出处
《中国生化药物杂志》
CAS
CSCD
北大核心
2011年第2期92-94,98,共4页
Chinese Journal of Biochemical Pharmaceutics
基金
广西科学研究与技术开发计划项目基金资助(桂科攻0424008-2F)
广西医科大学博士基金资助
关键词
菲牛蛭
克隆
序列分析
Hirudinaria manillensis
clone
sequence analysis
