| 阳帅，胡华，赵强，龚邵新，贺荣芳，张小丽，贺修胜.鼻咽癌相关新基因NPCEDRG突变型重组表达载体的构建及生物信息学分析.[J].中南医学科学杂志.,2013,41(6):551-554. |
| 鼻咽癌相关新基因NPCEDRG突变型重组表达载体的构建及生物信息学分析 |
| Construction of Eukaryotic Expression Recombinant of MutationalNPCEDRG and Its Bioinformatics Analysis |
| 投稿时间：2013-05-15 |
| DOI： |
| 中文关键词: NPCEDRG 聚合酶链式反应 随机突变 |
| 英文关键词:NPCEDRG polymerase chain reaction （PCR） random mutation |
| 基金项目:国家自然科学基金课题（81172575）资助. |
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| 中文摘要: |
| 目的构建带His标签的突变型及野生型pcDNATM3.1/myc-HisB/NPCEDRG重组表达载体。方法以pcDNA3.1-NPCEDRG重组质粒为模板，采用聚合酶链式反应（PCR）技术扩增NPCEDRG基因编码区，利用PCR过程中的碱基错配，随机突变产生NPCEDRG的突变体，分别将野生型及突变型NPCEDRG基因编码区cDNA插入pcDNATM3.1/myc-HisB真核表达载体，双酶切鉴定，测序验证，生物信息学方法进行蛋白质结构预测。结果 |
| 英文摘要: |
| ObjectiveTo construct two eukaryotic expression vectors of mutational and wild NPCEDRG by Random Mutation.MethodsThe ORF of NPCEDRG gene was amplified from the recombinant plasmid of pcDNA3.1-NPCEDRG by polymerase chain reaction （PCR）,and mutational NPCEDRG was procured by Random Mutation.The ORFs cDNAs of the wild NPCEDRG gene and its mutant were recombined respectively into the pcDNATM3.1/myc-HisB vector with two tags of myc and 6×His following the target genes.Then the products were transferred into E.coli DH5α,and the positive clones were screened after being identified with restrictive enzymes and sequence analysis.ResultsThe 519 bp target segments of mutational and wild NPCEDRG were obtained successfully in digestion products.The DNA sequence of wild NPCEDRG was consistent with the cDNA sequence of NPCEDRG from GeneBank.And the sequence of the mutational NPCEDRG revealed two altered codons,T260-C260 and T287-C287,which resulted in amino acid exchanges V73-A73 and M82-T82.ConclusionTwo recombinant expression vectors of mutational or wild NPCEDRG gene were constructed successfully. |
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