摘要
为了研究旋毛虫组织蛋白酶B的功能,对GenBank中旋毛虫基因组数据库和EST数据库进行检索,获得旋毛虫组织蛋白酶B的cDNA序列并设计引物。以旋毛虫肌幼虫总RNA为模板,进行RT-PCR,将PCR产物克隆到pMD18-T载体后测序并进行生物信息学分析。结果表明,成功克隆到旋毛虫组织蛋白酶B基因(TsCB1)的cDNA序列,TsCB1cDNA含有1个由1 449个核苷酸组成的完整的开放阅读框,编码482个氨基酸残基组成的多肽,蛋白质分子质量理论值为55.1ku,理论等电点为7.66。TsCB1第1~35位氨基酸残基为信号肽序列,有2个潜在的N-糖基化位点,具有1个生长调节素B结构域和半胱氨酸蛋白酶结构域,半胱氨酸残基活性位点被丝氨酸残基所替换,同源性分析表明与其他线虫组织蛋白酶B的一致性在60%左右。
In order to study the function of cathepsin B of Trichinella spiralis ,primers derived from T. spiralis genome and EST database in GenBank were designed and the open reading frame(ORF) se- quence of cathepsin B(TsCB1) was cloned by RT-PCR from T. spiralis muscle larvae,and bioinformatics analysis was performed. The results indicated that the TsCB1 cDNA sequence contained an ORF of 1 449 nucleotides and the deduced protein consisted of 482 amino acids with a theoretical molecular weight of 55.1 ku and an isoelectric point of 7.66. The deduced protein had a signal peptide sequence between amino acids 1 and 35,2 N-glycosylation sites,1 somatomedin B-2 domain and 1 cysteine protease domain with the cysteine active site changed to serine. TsCB1 protein sequence showed about 60% identity with cathepsin Bs from other parasitic nematodes.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2013年第7期661-667,共7页
Chinese Veterinary Science
基金
国家自然科学基金资助项目(31272555)
甘肃省创新研究群体计划项目(1210RJIA006)
关键词
旋毛虫
组织蛋白酶B
克隆
生物信息学分析
Trichinella spiralis
cathepsin t3
cloning
bioinformatics analysis