湖羊Sp1基因CDS区克隆及其对颗粒细胞增殖和凋亡的影响

doi:10.11843/j.issn.0366-6964.2017.11.011

Abstract

Abstract:

The aim of this study was to explore the sequence characteristics of Sp1 gene of Hu sheep and its effect on proliferation and apoptosis in granulosa cells. The CDS region of Sp1 gene was obtained by cloning and sequencing, and was analyzed by bioinformatics methods. Phylogenetic tree was constructed based on Sp1 amino acid sequence through NJ method. The expression profile of Sp1 was detected by RT-PCR. The Sp1 overexpression vector of Hu sheep was constructed by cloning, the activity of Caspase-3 and CCK-8 value was detected by correspoding kits; and the apoptosis rate of human ovarian granulosa cells (KGN) was detect by the flow cytometry. The results showed that the length of Sp1 CDS region was 2 340 bp and encoded 799 amino acid residues, which were 94.28% and 95.75% homology with the nucleotide sequence and amino acid sequence of human, respectively; Hu sheep had closer relationship with cow; Sp1 gene was widely expressed in many tissues of Hu sheep such as ovary, granulosa cells, and so on. Sp1 overexpression vector was constructed and transfected KGN cells successfully; the activity of Caspase-3 in the experimental group was remarkably higher than that in the control group (P<0.05) and CCK-8 value was just the reverse, which suggesting that overexpression of Sp1 gene inhibited the proliferation of KGN cells(P<0.05) and promoted the apoptosis of KGN cells (P<0.05).The Sp1 gene of Hu sheep was highly conserved in mammals, it could inhibit the proliferation of KGN and lead to its apoptosis.

CLC Number:

S826.2

YAO Yi-long, LI Yin-xia, REHEMAN Anwaier, ZHANG Jun, MENG Chun-hua, WANG Hui-li, QIAN Yong, CAO Shao-xian. Cloning of Sp1 Gene CDS Region of Hu Sheep and Its Effect on Proliferation and Apoptosis of Granulosa Cells[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(11): 2098-2106.

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Cloning of Sp1 Gene CDS Region of Hu Sheep and Its Effect on Proliferation and Apoptosis of Granulosa Cells

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