Research Article

Copy number and integration sites in growth hormone transgenic goats

Published: March 20, 2015
Genet. Mol. Res. 14 (1) : 2006-2014 DOI: https://doi.org/10.4238/2015.March.20.10
Cite this Article:
(2015). Copy number and integration sites in growth hormone transgenic goats. Genet. Mol. Res. 14(1): gmr4426. https://doi.org/10.4238/2015.March.20.10
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Abstract

Transgenic goats have been utilized for years to produce valuable protein. However, when transgenic goats are produced by random integration of inserted genes into cells, the copy number and integration sites of these genes in the goat genome are typically indefinite. Most polymerase chain reaction (PCR)-based methods that have been utilized to determine copy number and integration sites of inserted genes in the genome require complicated manipulations. In this study, we used quantitative real-time PCR and thermal asymmetric interlaced-PCR to determine copy number and integration sites of the inserted genes, respectively. Copies of transgenic goat lines GHcd-2 and GHcd-7 were 12.95 ± 0.18 and 12.24 ± 1.12, respectively. Two integration sites, located in chromosomes 3 and 11 and referred to as tg1 and tg2, were identified by thermal asymmetric interlaced-PCR. Junction PCR was then performed to confirm the integration sites of growth hormone transgenic goats. Transgenic copy number and integration sites were determined, which will be useful for determining the relationship between the growth hormone expression, copy number, and integration sites.

Transgenic goats have been utilized for years to produce valuable protein. However, when transgenic goats are produced by random integration of inserted genes into cells, the copy number and integration sites of these genes in the goat genome are typically indefinite. Most polymerase chain reaction (PCR)-based methods that have been utilized to determine copy number and integration sites of inserted genes in the genome require complicated manipulations. In this study, we used quantitative real-time PCR and thermal asymmetric interlaced-PCR to determine copy number and integration sites of the inserted genes, respectively. Copies of transgenic goat lines GHcd-2 and GHcd-7 were 12.95 ± 0.18 and 12.24 ± 1.12, respectively. Two integration sites, located in chromosomes 3 and 11 and referred to as tg1 and tg2, were identified by thermal asymmetric interlaced-PCR. Junction PCR was then performed to confirm the integration sites of growth hormone transgenic goats. Transgenic copy number and integration sites were determined, which will be useful for determining the relationship between the growth hormone expression, copy number, and integration sites.

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