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

Lin,Jian; Zhang,Qiang; Zhu,Li  Q.; Yu,Qing  H.; Yang,Qian

doi:10.3892/ijmm.2014.1841

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

Authors:
- Jian Lin
- Qiang Zhang
- Li Q. Zhu
- Qing H. Yu
- Qian Yang
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Affiliations: Key Laboratory of Animal Physiology and Biochemistry, Ministry of Agriculture, Nanjing Agricultural University, Nanjing, Jiangsu 210095, P.R. China
Published online on: July 9, 2014 https://doi.org/10.3892/ijmm.2014.1841
Pages: 900-910

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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.

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