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| The Relationship Between Exogenous DsRed Gene Copy Number and Exogenous Gene Expression in Transgenic Arbas Cashmere Goats (Capra hircus) |
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Abstract Exogenous genes mostly random integrated in the genome of transgenic animals which generated by somatic cell cloning and embryo transplantation. The copy number of exogenous gene is one of the important factors which affect the expression level of exogenous gene and genetic stability in transgenic animals. As the identification index of genetic stability of transgenic animals, it is necessary to detect the copy number of the exogenous gene after the survival of transgenic animals. In order to test whether there is relationship between the copies of exogenous red fluorescence protein from Discosoma sp. (DsRed) gene, and its expression level in the transgenic Arbas cashmere goat (Capra hircus) genomic, we used the absolute qRT-PCR method which was more efficient highly sensitive and accurate than the traditional method of Southern blot to identify the copy number of exogenous gene to test the copy number of 3 different fragments in exogenous DsRed gene in 6 transgenic cashmere goats. The results showed that the copy number of Cytomegalovirus (CMV) promoter ranged from 2.80±0.38 to 13.68±0.16, DsRed gene from 1.34±0.04 to 11.84±0.02, joint of CMV promoter and DsRed gene from 1.58±0.04 to 19.22±0.38. The expression of exogenous DsRed gene mRNA in 6 transgenic cashmere goats was detected by relative qRT-PCR method. The exogenous gene in 3 different fragments of copy number data respectively using SPSS software for correlation analysis, the correlation coefficient P value was, CMV promoter, P=0.763; DsRed gene, P=0.665; joint of CMV promoter and DsRed gene: P=0.803. The P values of the 3 groups were all greater than 0.05, The results showed that in our detection of 6 transgenic Arbas cashmere goats in exogenous DsRed gene expression and the copy number of the exogenous gene had no significant correlation at 0.05 level. In addition, the reliability of the qRT-PCR method in the study of the copy number of exogenous genes in large transgenic livestock was verified: With good reproducibility and controllable error, it can be the preferred method to detect the copy number of exogenous gene in transgenic animals.
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Received: 16 November 2015
Published: 06 May 2016
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