| 卢金凤,梁高峰,梁艳,黄倢.中国对虾Rab7全长cDNA的克隆及原核表达.渔业科学进展,2012,33(6):66-73 |
| 中国对虾Rab7全长cDNA的克隆及原核表达 |
| cDNA cloning and prokaryotic expression of Rab7 from shrimp Fenneropenaeus chinensis |
| 投稿时间：2012-02-28 修订日期：2012-04-25 |
| DOI： |
| 中文关键词: 中国对虾 fcrab7 克隆 原核表达 |
| 英文关键词: Fenneropenaeus chinensis fcrab7 RACE prokaryotic expression |
| 基金项目:公益性行业（农业）科研专项经费项目（201103034）、国家虾现代产业技术体系(nycytx 46) |
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| 中文摘要: |
| 本研究采用RACE方法克隆了中国对虾Fenneropenaeus chinensis Rab 7基因（fcrab7）全长cDNA (GenBank：JF742050）。生物信息学分析显示，fcrab7的开放阅读框615bp，编码205个氨基酸，分子量23.2kD。FcRab7的氨基酸序列具有Rab蛋白家族的共同特征，与凡纳滨对虾和斑节对虾同源蛋白的同源性为100%。构建重组表达载体pBAD/gIIIA-fcrab7转化到大肠杆菌E. coli进行诱导表达，诱导产物经SDS-PAGE检测和Western blot验证。本研究结果为进一步研究WSSV与FcRab7的相互作用提供了基础。 |
| 英文摘要: |
| Rab proteins are the largest subfamily in small GTP binding proteins, which play important roles in vesicle formation, transportation, adhesion, anchoring,and fusion. As a member of this family, Rab7 protein is in charge of the delivery of internalised materials into degradative compartments and the acquisition of lysosomal hydrolases. Evidence from the previous studies showed that VP28 of white spot syndrome virus (WSSV) can bind with Rab7 in Penaeus monodon (PmRab7). By rapid amplification of cDNA ends (RACE) method, the full length gene of Fenneropenaeus chinensis Rab7 gene (fcrab7) was obtained. The total cDNA consists of a 615bp open reading frame (ORF), which encoded 205 amino acids with a predicted molecular mass of 23.2 kDa . The deduced amino acid sequence contained many conserved motifs and regions, and the entire protein has 100% homology with Rab7 of Litopenaeus vannamei and P. monodon. The recombined vector of pBAD/gIIIA-fcrab7 was constructed and transformed into E. coli. The fusion protein was successfully expressed by induction, which was tested by SDS-PAGE and Western blot analysis. These results laid a basis for further studies on interaction between WSSV and FcRab7. |
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