Porcine Fetal Fibroblast Culture and Sex Identification of Banna Mini-pig Inbred Line

Biotechnology Bulletin ›› 2013, Vol. 0 ›› Issue (5): 116-120.

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Porcine Fetal Fibroblast Culture and Sex Identification of Banna Mini-pig Inbred Line

Xin Jige^{1, 2}, Xiao Jing², Zha Xingqin^{1, 2}, Cheng Wenmin^{1, 2}, Qing Yubo^{1, 2}, Pan Weirong^{1, 2}, Zeng Yangzhi², Wei Hongjiang^{1, 2}

(1. Animal Science and Technology College，Yunnan Agricultural University，Kunming 650201；2. Key Laboratory of Banna Mini-pig Inbred Line of Yunnan Province，Kunming 650201)

Received:2012-10-16 Revised:2013-05-24 Online:2013-05-24 Published:2013-05-24

Abstract

Abstract: The present study was designed to culture fetal fibroblast of Banna mini-pig inbred line and built sex identification method. Experiments were conducted to isolate fetal fibroblast of Banna mini-pig inbred line from embryos by collagenase digestion and identify their sex using multiplex PCR through optimizing conditions, in which two pairs of primer were designed relatively according to specific fragments in SRY gene and β-actin gene (as internal control). The results demonstrated that it could successfully isolate fetal fibroblast of Banna mini-pig inbred line and the embryo of number 2, 3 were male that could amplify specific DNA sequences of SRY gene (309 bp) and reference gene (132 bp) by multiplex PCR, and the number 1, 4, 5, 6, 7 were female with only reference gene. The PCR products were sequenced and aligned with SRY gene sequence published in NCBI. It shared 99% homology and this made to further verify the validity of results. The study provided a reliable technical basis for the culture of special sex fetal fibroblast in transgene and gene knock-out research.

Key words: Banna mini-pig inbred line, Fetal fibroblast, SRY gene

Xin Jige, Xiao Jing, Zha Xingqin, Cheng Wenmin, Qing Yubo, Pan Weirong, Zeng Yangzhi, Wei Hongjiang. Porcine Fetal Fibroblast Culture and Sex Identification of Banna Mini-pig Inbred Line[J]. Biotechnology Bulletin, 2013, 0(5): 116-120.

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Porcine Fetal Fibroblast Culture and Sex Identification of Banna Mini-pig Inbred Line

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