| 曹山，饶国栋，魏弘宜，蒋湘宁，陆海.毛白杨4CL-like基因的克隆与原核表达分析[J].广东农业科学,2013,40(9):140-142 |
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毛白杨4CL-like基因的克隆与原核表达分析 |
| Cloning and prokaryotic expression of Populus tomentosa Carr. 4CL-like gene |
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| DOI： |
| 中文关键词: 毛白杨 4CL-like基因 克隆 原核表达 |
| 英文关键词: Populus tomentosa Carr. 4CL-like gene clone prokaryotic expression |
| 基金项目:国家自然科学基金(31170574) |
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| 摘要点击次数: 1381 |
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| 中文摘要: |
| 为得到毛白杨4CL-like基因并对其原核表达特性进行分析，利用毛白杨叶片的总RNA反转录得到的总cDNA为模板，利用PCR技术克隆得到毛白杨4CL-like基因（登录号：XP_002315339.1）。利用MEGA软件对4CL-like基因及其他9个来自不同物种的4CL基因进行进化树分析。最后通过构建pET30-4CL-like原核表达载体对该基因进行原核表达分析。结果显示，克隆得到的4CL-like基因CDS区全长为1758bp，编码585个氨基酸。系统进化分析表明，该基因与葡萄中的4CL-like5有很高的同源性。另外，SDS-PAGE分析表明，4CL-like基因在大肠杆菌BL21中成功表达，所表达融合蛋白的分子量约为60 ku。 |
| 英文摘要: |
| In order to clone and analyze the prokaryotic expression of 4CL-like gene from Populus tomentosa Carr., total RNA were isolated, and 4CL-like gene were cloned by PCR. Phylogenetic analysis of the 4CL-like gene compared with other nine 4CL genes form different species with MEGA. Then it was over -expressed in prokaryotic systems. The expression construct pET30 -4CL -like was preformed. Based on the results, the bioinformational analysis showed that the CDS region of the nucleotide sequence was 1 758 bp, which could code 585 predicted aminoacids. Phylogenetic analysis showed that this gene was most similar to 4CL-like5 in Vitis vinifera. The SDS-PAGE analysis showed that the 4CL-like gene was transformed into E. coli BL21 and expressed successfully, fusion protein with molecular weight of 60 ku was successfully expressed in transformant. |
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