人甘露糖结合凝集素相关丝氨酸蛋白激酶-2 C端基因的克隆与鉴定

J4 ›› 2011, Vol. 37 ›› Issue (1): 51-55.

人甘露糖结合凝集素相关丝氨酸蛋白激酶-2 C端基因的克隆与鉴定

刘洁, 贾天军, 刘雪晴, 贾晓辉, 翟雪琼

（河北北方学院医学检验学院|河北张家口 075000）

收稿日期:2010-04-19 出版日期:2011-01-28 发布日期:2011-01-28
通讯作者: 贾天军 E-mail:jiatianjun@yahoo.com
作者简介:刘洁(1977-)|女|河北省阜城县人|生物化学与分子生物学硕士|主要从事天然免疫分子的研究。
基金资助:
河北省科技厅自然科学基金资助课题(C2006000875)

Cloning and identification of C-terminal gene encoding human mannose-binding lectin associated serine protein kinase-2

LIU Ji, JIA Tian-Jun, LIU Xue-Qing, JIA Xiao-Hui, ZHAI Xue-Qiong

(College of Medical Laboratory,HeBei NorthUniversity,Zhangjiakou,075000)

Received:2010-04-19 Online:2011-01-28 Published:2011-01-28

摘要/Abstract

摘要：

目的：获得人甘露糖结合凝集素相关丝氨酸蛋白激酶-2（MASP-2）C端编码基因，并表达人MASP-2 C端片段，为制备单克隆抗体及应用于临床相关疾病的检测奠定分子学基础。方法：采用RT-PCR技术从人胎肝组织总RNA中扩增人MASP-2 C端的cDNA，克隆入pGEX-6p-2表达载体，酶切图谱分析和序列分析鉴定。结果：获得编码人MASP-2 C端片段的cDNA，并且与pGEX-6p-2载体连接，转化大肠杆菌XL1-blue，成功构建人MASP-2 C端片段cDNA的重组克隆。重组质粒酶切图谱分析和DNA测序分析，人MASP-2 C端片段cDNA的重组克隆与GenBank中人MASP-2 C端的cDNA序列一致。结论：成功克隆了人MASP-2 C端片段cDNA，并在大肠杆菌中表达人MASP-2 C端多肽片段

Abstract:

bstract:Objective
To obain the C terminal gene encoding human mannose-binding lectin associated serine protein kinase-2(MASP-2),and express human MASP-2 C-terminal fragment and clinical application in E.coli and lay foundation for preparation of monoclonal antibodies and clinical application in relevant diseases. Methods The cDNA of human MASP-2 C-terminal was amplified from total RNA extracted from human fetal liver tissue with RT-PCR,and was cloned into pGEX-6p-2 vector and identified by restrictive digestion analysis and sequencing. Results The cDNA encoding human MASP-2 C-terminal was isolated,linked with pGEX-6p-2 vector and transformed into E.coli XL1-blue. The restriction map analysis and sequencing results were consistent with the sequence of MASP-2 C-terminal cDNA that reported in GenBank. Conclusion The human MASP-2 C-terminal cDNA is successfully cloned and expresses in E.coli.

Key words: mannose-binding lectin associated serine protein kinase-2 C;reverse transcriptase polymerase chain reaction;cloning,molecular;gene expression

中图分类号:

R349.6

刘洁, 贾天军, 刘雪晴, 贾晓辉, 翟雪琼. 人甘露糖结合凝集素相关丝氨酸蛋白激酶-2 C端基因的克隆与鉴定[J]. J4, 2011, 37(1): 51-55.

LIU Ji, JIA Tian-Jun, LIU Xue-Qing, JIA Xiao-Hui, ZHAI Xue-Qiong. Cloning and identification of C-terminal gene encoding human mannose-binding lectin associated serine protein kinase-2[J]. J4, 2011, 37(1): 51-55.