小鼠Lrh-1基因与排卵数间相关性分析
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王利红(1975-),女,研究方向:动物遗传繁育。E-mail:wanglihong345@yahoo.com.cn

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江苏畜牧兽医职业技术学院课题(HX200906);江苏省自然科学基金项目(BK2010356)。


Correlation analysis of Lrh-1 gene and ovulation number in mice
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    摘要:

    目的探讨肝受体类似物-1(liver receptor homolog-1, Lrh-1; NR5A2)基因序列差异与小鼠排卵数性状间关系。方法根据GenBank中NM_001159769序列设计了5对引物扩增Lrh-1基因CDS区,用单链构象多态性 (Single-strand conformation polymorphism,SSCP)分析小鼠Lrh-1基因序列差异,分析Lrh-1基因与小鼠排卵数间关系。结果①在5对引物中只有引物P2和P5扩增产物存在多态性,引物P2扩增产物存在两种基因型AA和AB型,其中AB型发生(C674A)突变,这种突变导致编码的140位氨基酸由谷氨酰胺变为赖氨酸。②引物P5 扩增产物存在三种多态类型SSCP-1、SSCP-2和SSCP-3,SSCP-1和SSCP-3型核酸序列比SSCP-2型少25个碱基,SSCP-3型除缺失区段外,其他区域碱基序列与SSCP-2型有5处碱基突变,而与SSCP-1型核酸序列有34处碱基差异,经BLAST分析,SSCP-2型与NM_001159769序列相似,SSCP-1型与NM_001159769序列相差较多,而与NG_012313.1序列相似,通过氨基酸序列比对分析,SSCP-3型1652处的A→G的突变导致氨基酸由丙氨酸变为苏氨酸,1678位G→C的突变使原密码子TAC(酪氨酸Y)变为TAG(终止子)。③引物P2 AA型(27.27±8.52)与AB型小鼠的平均排卵数(25.92±11.73)间差异无显著性(P>0.05);引物P5 SSCP-2型平均排卵数(35.00±14.58)显著高于SSCP-3型平均排卵数(19.50±7.94)(P<0.05),SSCP-1型(26.2±8.18)与SSCP-2型、SSCP-3型平均排卵数间差异无显著性(P>0.05)。结论明确了Lrh-1基因序列差异与小鼠排卵数性状间关系,为深入研究Lrh-1基因对动物生殖调控机理提供依据。

    Abstract:

    Objective To explore the correlation of liver receptor homolog-1(Lrh-1; NR5A2) gene nucleotide sequence and ovulation number in mice. MethodsFive primers were designed according to the sequence of NM_001159769 and amplified the coding sequence of Lrh-1 gene. Single-strand conformation polymorphism was used to analyze the correlation of difference in Lrh-1 nucleotide sequence and ovulation number. Results(1) There were polymorphisms in P2 and P5 PCR products. The P2 PCR products had two genotypes:AA and AB. The AB genotype had one base mutation of C674A, and the mutation led to amino acid change of Q140K. (2) The P5 PCR products had three SSCP-types:SSCP-1, SSCP-2 and SSCP-3. The nucleotide sequences of SSCP-1 and SSCP-3 had 25 base deletion than SSCP-2 type. Five base mutations between SSCP-3 and SSCP-2,4 base mutations between SSCP-3 and SSCP-1 were found. When BLAST in NCBI, the nucleotide sequence of SSCP-2 was like NM_001159769, but SSCP-1 sequence was like NG_012313.1. The base mutation of A1652G caused alanine changed to threonine and the base mutation of G1678C caused tyrosine changed to termination codon in peptide chain of SSCP-3 type. (3)There was no significant difference of average ovulation numbers between AA genotype (27.27±8.52) and AB genotype (25.92±11.73) in P2 PCR products (P>0.05). In P5 PCR products, the SSCP-2 type (35.00±14.58) had a significantly higher average ovulation number than SSCP-3 type’s (19.50±7.94) (P<0.05). ConclusionsDefinite correlations of Lrh-1 gene and ovulation number have been established in mice. The results will contribute to the research of Lrh-1 gene molecular mechanism and regulatory pathway in animal reproduction.

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王利红,张伟,高勤学,王子玉,张艳丽,王锋.小鼠Lrh-1基因与排卵数间相关性分析[J].中国实验动物学报,2012,(1):65~69.

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