[1]霍瑞清,白瑞霞.人CyclinD1在毕赤酵母中的表达[J].现代检验医学杂志,2016,31(05):81-83，87.[doi:10.3969/j.issn.1671-7414.2016.05.021]
　HUO Rui-qing,BAI Rui-xia.Expression of Human CyclinD1 in Pichia Pastoris[J].Journal of Modern Laboratory Medicine,2016,31(05):81-83，87.[doi:10.3969/j.issn.1671-7414.2016.05.021]
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人CyclinD1在毕赤酵母中的表达()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第31卷
期数:
2016年05期
页码:
81-83，87
栏目:
论著
出版日期:
2016-10-21

文章信息/Info

Title:
Expression of Human CyclinD1 in Pichia Pastoris
文章编号:
1671-7414(2016)05-081-04
作者:
霍瑞清; 白瑞霞
内蒙古自治区人民医院检验科,呼和浩特 010017
Author(s):
HUO Rui-qing; BAI Rui-xia
Department of Clinical Laboratory,Inner Mongolia People's Hospital,Hohhot 010017,China
关键词:
Cyclin D1;  克隆;  毕赤酵母
分类号:
R735.7; R730.43
DOI:
10.3969/j.issn.1671-7414.2016.05.021
文献标志码:
A
摘要:
目的 克隆人肝癌组织中的Cyclin D1基因,并在毕赤酵母中表达Cyclin D1蛋白。方法 从人肝癌组织中提取RNA,逆转录PCR扩增获得Cyclin D1 cDNA,将PCR产物进行TA克隆和DNA序列分析; 阳性TA克隆Cyclin D1片段再亚克隆入毕赤酵母表达载体Ppic9k,甲醇诱导表达。结果 逆转录PCR扩增得到约483 bp的DNA片段,TA克隆和DNA序列分析结果显示重组片段为人的CyclinD1基因,Cyclin D1 cDNA亚克隆入Ppic9k载体的NotI和EcoRI位点之间; 甲醇诱导得到分子量约36KD的蛋白。结论 成功克隆人肝癌组织中的Cyclin D1基因,并且在毕赤酵母中成功表达。
Abstract:
Objective To obtain the Cyclin D1,the Cyclin D1 gene was cloned and expressed in Pichia pastoris.Methods The total RNA was extracted from liver cancer tissue.The Cyclin D1 cDNA wasobtained by RT-PCR.The Cyclin D1 cDNA was sequenced.And sub-clonedto the Ppic9k,the Pichia pastoris expressed was used to obtain the Cyclin D1.Results The 483 bp Cyclin D1 cDNA was obtained.And the sequence ofCyclin D1 was corrected.The 36KD Cyclin D1 was obtained by Pichia pastoris expression.Conclusion The Cyclin D1 cDNA was cloned and Cyclin D1 was expressed in Pichia pastoris.

参考文献/References:

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备注/Memo

备注/Memo:
基金项目:内蒙古自治区高等学校科学研究项目(NJZY13434)。
作者简介:霍瑞清(1966-),女,大专,副主任护师,研究方向:微生物感染,Tel:18047191527,E-mail:huoruiqing@126.com。
通讯作者:白瑞霞(1973-),女,博士,副教授,研究方向:分子生物学,Tel:18047193127,E-mail: yxybrx@126.com。
更新日期/Last Update: 2016-10-30