Song Shaozheng, Zhang Min, Li Dan, Chen Chaojun, Yu Kangying, Zhang Ting, Zhou Mingming, Cheng Yong
School of Nursing, Taihu University of Wuxi, Wuxi 214000, Jiangsu, China.
Jiangsu Provincial Research Center for Animal Transgenesis and Biopharming, College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, Jiangsu, China.
Sheng Wu Gong Cheng Xue Bao. 2020 Jan 25;36(1):67-76. doi: 10.13345/j.cjb.190173.
The genetic background such as copy number, integration site and chromosome karyotype of exogenous genes of transgenic animals obtained by random integration is still unclear. There may be some problems such as silent integration, invalid integration, toxic integration and unpredictable expression level of exogenous genes. In this study, six primary (F0) and their corresponding offspring (F1) of human lactoferrin (hLF) transgenic goats were selected as the research objects, and blood samples were collected from jugular vein and DNA were extracted. The genetic background and expression level of exogenous genes were studied by chromosome karyotype analysis, real-time quantitative PCR (qPCR), ELISA and Western blotting. The chromosomes of six F0 transgenic goats had no obvious morphological variation, number change and other abnormalities. The relative copy number was different (2-16) and could be steadily inherited to the next generation. The copy number of F0 and F1 hLF gene was the same. The highest expression level of hLF was 1.12 g/L in F1 transgenic goats (L3-1, 8 copies). The results proved that the integrated exogenous genes could steadily inherit the next generation, and did not cause obstacles to the growth and development of transgenic goat individuals. Moreover, there was no obvious correlation between the number of copies and the expression level of hLF. This laid a foundation for the new varieties cultivation of transgenic goats and other transgenic animals, and analysis of genetic background.
