[1]谭虎,杨天德,粟永萍,等.修饰GLP2分泌性表达载体的构建及促增殖活性测定的实验研究[J].陆军军医大学学报(原第三军医大学学报),2007,29(17):1650-1653.
 TAN Hu,YANG Tian-de,SU Yong-ping,et al.Construction of secretary expression vector of mutation chimeric GLP2 and detection of its proliferation-promoting activity[J].J Amry Med Univ (J Third Mil Med Univ),2007,29(17):1650-1653.
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修饰GLP2分泌性表达载体的构建及促增殖活性测定的实验研究(/HTML )
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
29卷
期数:
2007年第17期
页码:
1650-1653
栏目:
论著
出版日期:
2007-09-15

文章信息/Info

Title:
Construction of secretary expression vector of mutation chimeric GLP2 and detection of its proliferation-promoting activity
作者:
谭虎杨天德粟永萍艾国平黄岚陶军刘晓宏楼淑芬
第三军医大学军事预防医学院防原医学教研室,全军复合伤研究所,创伤、烧伤与复合伤国家重点实验室;第三军医大学新桥医院麻醉科;第三军医大学新桥医院全军心血管病研究所
Author(s):
TAN Hu YANG Tian-de SU Yong-ping  AI Guo-ping  HUANG Lan TAO Jun LIU Xiao-hong LOU Shu-fen
State Key Laboratory of Trauma, Burns and Combined Injury, Institute of Combined Injury, College of Preventive Medicine, Chongqing 400038,Department of Anesthesiology,Department of Cardiovascular Diseases, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China
关键词:
胰高血糖素样肽-2基因克隆PCR重叠延伸技术肠上皮细胞增殖
Keywords:
glucagon-like peptid-2 gene clone splicing by overlap extension intestine epithelium cell proliferation
分类号:
R329.25;R394-33;R394.2
文献标志码:
A
摘要:
目的  对胰高血糖素样肽-2(glucagon-like peptid-2, GLP-2)进行基因修饰,观察其促细胞增殖活性。  方法  通过RT-PCR方法获得含GLP2编码序列的高血糖素原cDNA片段;通过PCR重叠延伸技术得到GLP2信号肽及成熟肽的嵌合分子,并将编码GLP2成熟肽N-末端第二位丙氨酸的序列(GCT)突变为编码甘氨酸的序列(GGT);将其插入真核表达质粒pVITRO3,转染至肠上皮细胞(Caco-2),观察其对细胞增殖活性的影响。  结果  克隆的修饰GLP2基因与国外报道的核苷酸序列及其编码的氨基酸序列完全一致,并成功拼接GLP2的信号肽及成熟肽;且加入pVITRO3-GLP2-Caco-2培养上清的细胞生长明显快于对照组。  结论  PCR重叠延伸技术适用于分泌性载体构建。构建的修饰GLP2表达系统所表达的蛋白具有促进肠上皮细胞增殖的作用。
Abstract:
Objective    To construct the secretary expression vector of mutation chimeric GLP2 by clone technology and detect its effect on  cell proliferation.     Methods    The proglucagon cDNA was generated by the reverse transcription. The PCR product of signal and mature peptides of GLP2 was linked by SOE (splicing by overlap extension) and the gene, which coded the second amino acid in the N-terminal of GLP2, was mutated from GCT to GGT by PCR. Then the chimeric GLP2 cDNA was cloned into pVITRO3 vector and the recombinant plasmid was transfected to Caco-2. The cell proliferation was detected by MTT.     Results    The sequences of signal peptide and mature peptide of GLP2 were connected successfully and the mature peptide sequence was mutated. MTT and 3H-TdR showed that rGLP2 could accelerate the proliferation of Caco-2 cell significantly.     Conclusion    SOE is appropriate in establishing secretion type of expression vector and the secretary expression vector of mutation chimeric GLP2 has accelerating effect on cell proliferation.

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更新日期/Last Update: 2008-07-21