摘要
利用RT-PCR和RACE技术,从凡纳滨对虾(Litopenaeus vannamei)肝胰腺中克隆了DNaseⅠ基因的全长cDNA序列。该序列全长1614bp,包含1209bp的开放阅读框,编码一个含403个氨基酸的蛋白;5′非翻译区为116bp,3′非翻译区为289bp。实时定量PCR分析结果表明,DNaseⅠ基因在肝胰腺的表达量是其他器官表达量的16～162倍,表明凡纳滨对虾DNaseⅠ基因属于胰腺型表达。本研究还利用酶切重组构建原核表达载体,并在大肠杆菌(Escherichia coli)中成功表达出了有活性的重组DNaseⅠ蛋白。
The complete cDNA sequence of DNase I was isolated from Litopenaeus vannamei hcpatopancreas by RT-PCR and RACE. The full length eDNA of DNase I was 1 614 hp, consisting of a 5＇-noncoding region of 116 bp, a 3＇-noncoding region of 289 bp, and an open reading frame of 1 209 bp encoding a polypeptide of 403 amino acids. Real time quantitative PCR analysis revealed that the expression level of DNase I gene of L. vannamei in hepatopancreas was 16 - 162 folds higher than that in other 5 selected organs and DNase I gene of L. vannamei belonged to hepatopancreas type. Furthermore, a DNase I protein expression system based on Escherichia coli was constructed and functional recombinant DNase I protein was obtained successfully.
出处
《动物学杂志》
CAS
CSCD
北大核心
2010年第1期8-17,共10页
Chinese Journal of Zoology
基金
国家自然科学基金项目(No.30571430)
集美大学创新团队基金项目(No.2008A001)
