牦牛KDM1A基因克隆及其在不同发育时期睾丸中的表达规律

doi:10.11843/j.issn.0366-6964.2018.12.028

摘要/Abstract

摘要：

本研究旨在克隆牦牛组蛋白去甲基化酶1A（lysine-specific histone demethylase 1A，KDM1A）基因，检测其在牦牛不同组织及睾丸发育过程中的表达水平。采集4~5岁健康牦牛心、脾、肝、卵巢、肺、大脑、肾、子宫、大肠、睾丸和胃，提取各组织样的总RNA，另采集不同发育时期睾丸组织：胎牛（5~6月）、幼年时期（1~2岁）、性成熟时期（4~5岁），老年时期（9~10岁）。利用RT-PCR方法获取牦牛KDM1A的CDS区基因序列，并使用生物信息软件分析其结构和功能。采用实时荧光定量PCR（quantative real-time PCR，RT-qPCR）检测KDM1A在不同组织中的表达水平，并探究不同发育时期睾丸中KDM1A的表达规律。结果表明，本研究克隆获得牦牛KDM1A基因2 401 bp的cDNA序列。牦牛KDM1A核苷酸序列与野牦牛、黄牛的同源性较高，表明该基因在进化过程中较为保守。牦牛KDM1A基因CDS区为2 331 bp，编码776个氨基酸残基。KDM1A在各组织中均有表达，其中睾丸和肝中表达量最高。KDM1A mRNA的表达水平呈先上升（从胎牛~性成熟时期）再下降（性成熟~老年时期）的趋势。综上表明，本研究成功克隆了KDM1A基因，其在不同发育时期睾丸中表达规律不同，KDM1A基因可能参与牦牛睾丸的发育。

Abstract:

The aim of this study was to clone the lysine-specific histone demethylase 1A (KDM1A) gene, identify its expression in various tissues of yak, and to analyze the expression pattern in different growth periods of yak testis. The total RNA was extracted by collecting the heart, spleen, liver, ovary, lung, cerebrum, kidney, uterus, large intestine, testis and stomach from healthy yaks aged 4-5 years old.In addition,the testes at different developmental stages were collected including fetus (5-6-month-old), infancy (1-2-year-old), sexual maturity (4-5-year-old) and old age (9-10-year-old).RT-PCR was adopted for amplification of the complete CDS of KDM1A gene in yak.Meanwhile the structure and function of KDM1A gene in yak were analyzed by a series of bioinformatics softwares.Then the expression of KDM1A in different tissues and testes at different developmental stages were detected by RT-qPCR.The results showed that the KDM1A was obtained by cDNA cloning,the length of which was 2 401 bp.And the length of the KDM1A gene was 2 331 bp in CDS, encoding 776 amino acids.It was high homology identity to that of corresponding cDNA from bovine,which showed that the KDM1A gene was conservative in the process of evolution.The expression profile of KDM1A was wide in yak tissues,but there are some differences in various tissues.For example,the expression level was the highest in liver and testis.The expression level of KDM1A mRNA during testis development presented the tendency of going up firstly and going down secondly. The complete CDS of KDM1A gene was successfully cloned and the KDM1A had a significant different expression among the 4 periods of yak testis,which indicted that the KDM1A gene might play an important role in testis development of yak.

中图分类号:

S823.8⁺5.2

韩杰, 熊显荣, 王艳, 杨显英, 阿果约达, 黄向月, 李键. 牦牛KDM1A基因克隆及其在不同发育时期睾丸中的表达规律[J]. 畜牧兽医学报, 2018, 49(12): 2777-2785.

HAN Jie, XIONG Xian-rong, WANG Yan, YANG Xian-ying, A GUO Yue-da, HUANG Xiang-yue, LI Jian. Cloning of Yak KDM1A Gene and Its Expression Profile in Testis during Different Developmental Stages[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(12): 2777-2785.

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