转基因克隆牛外源基因整合位点的检测

doi:10.11843/j.issn.0366-6964.2014.04.007

畜牧兽医学报 ›› 2014, Vol. 45 ›› Issue (4): 553-558.doi: 10.11843/j.issn.0366-6964.2014.04.007

转基因克隆牛外源基因整合位点的检测

王敬姣¹，张明月¹，谭贝贝¹，张萃¹，李冬杰²，戴蕴平³，李宁³，李世杰^1*

(1.河北农业大学生命科学学院，保定 071001； 2.河北科技大学生物科学与工程学院，石家庄 050018；3.中国农业大学农业生物技术国家重点实验室，北京 100193)

收稿日期:2013-11-20 出版日期:2014-04-23 发布日期:2014-04-23
通讯作者: 李世杰，教授，E-mail：lishijie20005@163.com
作者简介:王敬姣(1987-)，女，河北保定人，硕士生，主要从事动物分子遗传方面的研究，E-mail:13931391932@163.com
基金资助:
国家自然科学基金(31372312)；河北省自然基金项目（C2011204001）

Integration Site Detection of Exogenous Gene in Transgenic Cloned Cattle

WANG Jing-jiao¹，ZHANG Ming-yue¹，TAN Bei-bei¹，ZHANG Cui¹，LI Dong-jie²，DAI Yun-ping³ ，LI Ning³ ，LI Shi-jie^1*

(1.College of Life Science，Agricultural University of Hebei，Baoding 071001， China;2.College of Life Science and Technology，Hebei University of Science and Technology，Shijiazhuang 050018， China; 3.State Key Laboratory for Agrobiotechnology，China Agricultural University，Beijing 100193， China)

Received:2013-11-20 Online:2014-04-23 Published:2014-04-23

摘要/Abstract

摘要：

确定外源基因的整合位点是获得转基因动物后的首要任务。本试验应用 TAIL-PCR得到了2头转入人溶菌酶基因克隆牛外源基因的整合位点，结果表明：2头转基因个体中外源基因均整合在受体基因组上,其中WS个体外源基因整合在牛24号染色体的基因组克隆(NW_003104566.1)中。YW个体中外源基因整合在牛16号染色体的基因组克隆(NW_003104440.1)中，整合位点位于LOC10030和RGL1基因之间。综上表明，在WS和YW个体中，外源基因的整合分别造成受体染色体238和228 bp核苷酸的缺失。4个整合片段（InS1~InS4）的末端均与拓扑异构酶I作用位点的共有序列相连。在WS个体中外源基因与染色体的整合连接处（InS1~InS3）存在1 nt的同源序列。在整合片段InS2、InS3、InS4中表现出共同的相似处，即均存在具有间隔的短的同源序列。

Abstract:

After produce the transgenic animals，the primary task was important to detect integration sites of these animals.In this study，the integration sites were successfully cloned in two transgenic SCNT cattles with hLYZ gene by TAIL-PCR.The results showed that in two SCNT transgenic cattles，the exogenous genes integrated to cattle chromosome.The exogenous gene in the WS individual integrated to genomic clones (NW_003104566.1) on bovine chromosome 24 and that in the YW individual integrated to genomic clones (NW_003104440.1) on bovine chromosome 16，which located between LOC10030 and RGL1 gene.The deletions of 238 and 228 bp host genome were detected in WS and YW respectively.The consensus sequence for mammalian Topo I ( topoisomerase-I ) cleavage sites were found at 4 integration sequences (InS1-InS4).At the junction of exogenous gene and host genome (InS1 and InS3) in WS，homologous sequences of 1 nt were found.InS 2，InS 3 and InS 4 showed similar homologous sequences at the junction.

中图分类号:

S823
Q78

王敬姣，张明月，谭贝贝，张萃，李冬杰，戴蕴平，李宁，李世杰. 转基因克隆牛外源基因整合位点的检测[J]. 畜牧兽医学报, 2014, 45(4): 553-558.

WANG Jing-jiao，ZHANG Ming-yue，TAN Bei-bei，ZHANG Cui，LI Dong-jie，DAI Yun-ping，LI Ning,LI Shi-jie. Integration Site Detection of Exogenous Gene in Transgenic Cloned Cattle[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(4): 553-558.

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转基因克隆牛外源基因整合位点的检测

Integration Site Detection of Exogenous Gene in Transgenic Cloned Cattle

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