| 兰旺仁,张子平,邹志华,王国栋,王艺磊.2010.凡纳滨对虾DNaseⅠ基因的克隆及原核表达.动物学杂志,45(1):8-17. |
| 凡纳滨对虾DNaseⅠ基因的克隆及原核表达 |
| Cloning and Prokaryotic Expression of DNaseⅠ from Litopenaeus vannamei |
| 投稿时间：2009-04-08 修订日期：2009-11-09 |
| DOI： |
| 中文关键词: DNaseⅠ RACE 原核表达 凡纳滨对虾 |
| 英文关键词:DNaseⅠ RACE Prokaryotic expression Litopenaeus vannamei |
| 基金项目: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) |
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| 摘要点击次数: 1949 |
| 全文下载次数: 2015 |
| 中文摘要: |
| 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。 |
| 英文摘要: |
| The complete cDNA sequence of DNaseⅠ was isolated from Litopenaeus vannamei hepatopancreas by RT-PCR and RACE.The full length cDNA of DNaseⅠ was 1 614 bp,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 DNaseⅠ gene of L.vannamei in hepatopancreas was 16-162 folds higher than that in other 5 selected organs and DNaseⅠ gene of L.vannamei belonged to hepatopancreas type.Furthermore,a DNaseⅠ protein expression system based on Escherichia coli was constructed and functional recombinant DNaseⅠ protein was obtained successfully. |
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