Abstract
Porcine endogenous retrovirus (PERV), which integrates as a provirus into the genome of pig cells, is an important biosafety issue in xenotransplantation. Screening and analyzing the presence and expression of PERV will provide essential parameters for assessing the biosafety of donor sources. In the present study, we investigated the prevalence of PERV in Diannan small-eared pigs, a unique closed colony that is distributed in southern Yunnan Province in southwestern China. PCR was performed to amplify env-A, env-B, env-C, pol, gag, and mtDNA in peripheral blood samples. The results revealed that PERV env-A, env-B, pol, and gag were detected in all individuals, but env-C was deficient in most pigs, suggesting that the main subtypes of PERVs in Diannan small-eared pigs are PERV-A and PERV-B. Furthermore, PERV pol and the porcine housekee** gene GAPDH were detected by RT-PCR in all peripheral blood samples, indicating that PERV had transcriptional activity. Finally, the consensus sequences of PERV-A and PERV-B were amplified and digested with KpnI and MboI. Interestingly, a total of seven digestion patterns were obtained, which is less than that observed in other pig breeds. The PCR products were cloned into the pUCm-T vector and sequenced. The results showed that all of the inserts were highly homologous to either PERV-A or PERV-B, and the ratios of PERV-A and PERV-B were 21.1% and 78.9%, respectively. These data suggest that Diannan small-eared pigs may be a candidate donor source for xenotransplantation.
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Acknowledgements
This work was supported by grants from the National Natural Science Foundation of China (No. 81560140). We are grateful for the samples provided by the Animal Experimental Center of Kunming Medical University.
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Ruhong Li and Gang Liu designed the study. Yunfei Zhang, **aowei **ng, and Peng Li performed the study. Yunfei Zhang, **aowei **ng, Lihua Huang, and Yong Wu analyzed the data and wrote the manuscript. All the authors have read and approved the final manuscript.
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This study was approved by the Experimental Animals Ethics Committee of the Third **angya Hospital, Central South University.
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Zhang, Y., **ng, X., Huang, L. et al. Screening pigs for xenotransplantation in China: investigation of porcine endogenous retrovirus in Diannan small-eared pigs. Virus Genes 56, 202–208 (2020). https://doi.org/10.1007/s11262-019-01722-7
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DOI: https://doi.org/10.1007/s11262-019-01722-7