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SS-CP-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。
Objective To explore the correlation of liver receptor homolog-1 （Lrh-1; NRSA2） gene nucleotide sequence and ovulation number in mice. Methods Five primers were designed according to the sequence of NM001159769 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 192 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 Q14OK. （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, 34 base mutations between SSCP-3 and SSCP-1 were found. When BLAST in NCBI, the nueleotide sequence of SSCP-2 was like NM_001159769, but SSCP-1 sequence was like NG_012313. I. 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 be-tweenAA 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）. Conclusions Definite 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.
出处
《中国实验动物学报》
CAS
CSCD
2012年第1期65-69,共5页
Acta Laboratorium Animalis Scientia Sinica
基金
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)
