宽体金线蛭WpE基因的克隆及生物信息学分析

Cloning and Bioinformatics Analysis of WpE Gene from Whitmania Pigra

目的:克隆宽体金线蛭(WpE)基因,并进行生物信息学分析,为进一步研究宽体金线蛭药用价值提供理论基础。方法:从宽体金线蛭唾液腺转录组数据库筛选到WpE基因转录本,通过PCR反应验证WpE基因cDNA全长序列,运用生物信息学分析WpE基因碱基序列性质及其对应氨基酸序列的特性。结果:宽体金线蛭WpE基因碱基序列全长258 bp,编码85个氨基酸,其中包含一个由16个氨基酸组成的信号肽,成熟蛋白理论相对分子质量为8.093 kDa,理论等电点(PI)为5.85,该蛋白为亲水性蛋白,属于马铃薯抑制剂Ⅰ家族。结论:成功克隆出宽体金线蛭WpE基因,分析其序列的结构特征,该基因编码的蛋白质含有马铃薯抑制剂Ⅰ家族保守活性位点Asp46,其结合环能与胰凝乳蛋白酶结合并使其失活,具有抗炎与抗凝活性。

Objective:To clone WpE gene from Whitmania pigra and perform the bioinformatics analysis,which lays a solid foundation for the further research on the medicinal value of W.pigra.Methods:The transcripts of WpE gene were screened from the saliary gland transcriptome database of W.pigra.The full-length cDNA sequence of WpE gene was verified by PCR reaction,and the properties of the base sequence and corresponding amino acid sequence of WpE gene were analyzed by bioinformatics.Results:The base sequence full-length of WpE gene was 258 bp,which encoded 85 amino acids,which includes signal peptide consisting of 16 amino acids;the mature protein′s theoretical molecular weight was 8.093 kDa,and its isoelectric point was 5.85,and the WpE protein was a hydrophilic protein,belonging to the potato inhibitorⅠfamilies.Conclusion:The WpE gene was successfully cloned and its structural characteristics were analyzed.The protein encoded by this gene contains Asp46,the conserved active site of potato inhibitorⅠfamily,and its binding ring can bind to chymotrypsin and inactivate it,showing anti-inflammatory and anticoagulant activities.