牦牛KDM4A基因克隆、组织表达谱及其在卵母细胞和颗粒细胞中的表达

doi:10.11843/j.issn.0366-6964.2018.02.008

Abstract

Abstract:

The aim of this study was to clarify the expression characteristics of yak KDM4A gene and its role in yak oocytes and granulosa cells. The primer was designed according to the published bovine (Bos taurus) KDM4A gene sequence on the GenBank, and the expression pattern of KDM4A was detected by RT-PCR in yak various tissues which included heart, liver, spleen, lung, kidney, small intestine, ovary, uterus, stomach, testicle and cerebrum.While the coding sequence of KDM4A gene was cloned by RT-PCR.The structure and function of KDM4A were analyzed by ExPASY software on line. The mRNA expression level of KDM4A in different periods of yak oocytes and granulosa cells were detected by qRT-PCR. Results showed that KDM4A gene of yak contained a 3 289 bp cDNA fragment, and had high homology with other mammals in nucleotide sequence through sequence alignment and phylogenetic tree, indicating that KDM4A was relatively conservative in the evolutionary process. The complete CDs of yak KDM4A gene was 3 201 bp, encoding 1 066 amino acids, and the molecular weight was 122.48 ku. The protein encoded by KDM4A gene in yak was an unstable, soluble and acidic protein without transmembrane regions and signal peptide. Random coil and α-helix were mainly in the secondary structure of KDM4A, consistent with the results of the tertiary structure analysis. KDM4A gene was expressed in various tissues of yak, and high abundance in ovary, spleen and testicle.The qRT-PCR results showed that the mRNA expression level in MⅡ phase granulosa cell was significantly higher than that in GV and MI phases (P<0.05). Meanwhile, the mRNA expression level of KDM4A was significantly higher in GV phase oocyte than that in MI and MⅡ phases (P<0.05). The complete CDs of KDM4A gene was successfully cloned and the KDM4A had a significant difference expression among the 3 periods of yak oocytes and granulosa cells, which indicated that KDM4A played role in the maturing process of yak oocytes and granulosa cells.

CLC Number:

S823.8⁺5.2

HAN Jie, XIONG Xian-rong, CAI Wen-yi, YANG Xian-ying, A GUO Yue-da, ZHANG Yan-hong, LI Jian. Cloning of KDM4A Gene and Its Expression in Different Tissues, Oocyte and Granulosa Cell of Yak[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(2): 291-299.

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Cloning of KDM4A Gene and Its Expression in Different Tissues, Oocyte and Granulosa Cell of Yak

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