ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2014, Vol. 45 ›› Issue (4): 553-558.doi: 10.11843/j.issn.0366-6964.2014.04.007

Previous Articles     Next Articles

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-jie1*   

  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

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.

CLC Number: