The copy number and integration site analysis of IGF-1 transgenic goat

Int J Mol Med. 2014 Sep;34(3):900-10. doi: 10.3892/ijmm.2014.1841. Epub 2014 Jul 9.

Abstract

Transgenic animals have been used previously to study gene function, produce important proteins, and generate models for the study of human diseases. As the number of transgenic species increases, reliable detection and molecular characterization of integration sites and copy number are crucial for confirming transgene expression and genetic stability, as well as for safety evaluation and to meet commercial demands. In this study, we generated four transgenic goats by somatic cell nuclear transfer (SCNT). After birth, the cloned goat contained transferred insulin-like growth factor I (IGF-1) gene was initially confirmed using a polymerase chain reaction (PCR)‑based method. The four cloned goats were identified as IGF-1 transgenic goats by southern blotting. The number of copies of the IGF-1 gene in each of the transgenic goats was determined. Additionally, four integration sites of the transgene in the transgenic goats with a modified thermal asymmetric interlaced (TAIL)-PCR method were identified. The four different integration sites were located on chromosomes 2, 11, 16 and 18. The present study identified the copy number and integration sites using quantitative PCR (qPCR) and TAIL-PCR, enabling the bio-safety evaluation of the transgenic goats.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Base Sequence
  • Blotting, Southern
  • Gene Dosage*
  • Goats / genetics*
  • Humans
  • Insulin-Like Growth Factor I / genetics*
  • Molecular Sequence Data
  • Mutagenesis, Insertional / genetics*
  • Polymerase Chain Reaction
  • Reference Standards

Substances

  • Insulin-Like Growth Factor I