Cloning and sequencing analysis of human PKCβⅡgene ORF

DUAN Lian; et al

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Cloning and sequencing analysis of human PKCβⅡgene ORF

更新时间：2024-01-17

- Cloning and sequencing analysis of human PKCβⅡgene ORF
- Journal of Chongqing Medical University Issue 1, Pages: 18-21(2008)
- 作者机构：
  
  1. 重庆医科大学附属第一医院内分泌科
  2. 重庆医科大学附属第一医院内分泌科,重庆,400016
- 作者简介：
- 基金信息：
- DOI：
  CLC： Q78
- Published：2008，
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[1]段炼,周波,李启富.人蛋白激酶CβⅡ基因开放读码框的克隆及序列分析[J].重庆医科大学学报,2008(01):18-21.

DUAN Lian, et al. Cloning and sequencing analysis of human PKCβⅡgene ORF. [J]. Journal of Chongqing Medical University (1):18-21(2008)
[1]段炼,周波,李启富.人蛋白激酶CβⅡ基因开放读码框的克隆及序列分析[J].重庆医科大学学报,2008(01):18-21. DOI：

DUAN Lian, et al. Cloning and sequencing analysis of human PKCβⅡgene ORF. [J]. Journal of Chongqing Medical University (1):18-21(2008) DOI：

摘要

目的:克隆人蛋白激酶CβⅡ基因（PRKCB1）的开放读码框（Open reading frame

ORF）

为其进一步的功能研究提供基础。方法:采用逆转录-巢式PCR法

从人脐静脉内皮细胞株中扩增出含PRKCB1基因ORF全长的片段

所得片段经加"A"处理并插入T载体。经蓝白筛选

用特异引物扩增阳性重组子

得到编码人蛋白激酶CβⅡ（Protein kinase CβⅡ

PKCβⅡ）基因的ORF

并与T载体相连

测序鉴定。结果:通过巢式RT-PCR及T/A克隆获取了PRKCB1基因的ORF

测序结果与GenBank报道序列一致。结论:成功克隆了PRKCB1基因的ORF

在技术路线上可以为某些难克隆的基因提供参考。

Abstract

Objective:To clone human PRKCB1 gene open reading frame(ORF) in order to further research on it’s function.Methods:RT-nested PCR method was adopted to the amplify the total length of human PRKCB1 ORF from human umbilical vein endothelial cell(HUVEC).T vector was inserted into the harvested fragment after a tail was added.With blue white screening

the gene ORF encoding human PKCβⅡ was obtained by special primers amplifying recombinant plasmids

and linked into T vector.Then the plasmid was identified by sequencing.Results:The human PRKCB1 ORF was amplified successfully with RT-nested PCR and T/A cloning

and the gene sequence was completely consistent with that reported in GenBank.Conclusion:Human PRKCB1 gene ORF was successfully cloned.The strategy of cloning may provide technical references for some genes hard to be cloned.

关键词

蛋白激酶Cβ基因聚合酶链反应T/A克隆序列分析

Keywords

PKCβ genePCRT/A cloningsequence analysis

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