利用单倍型分析在已知QTL内鉴定小鼠性发育候选基因
发布时间：2019-04-16 14:44
【摘要】：性发育是指个体性成熟并获得生殖能力的过程,受到多种因素的影响。性发育的启动时间是性发育过程中相当关键的一个性状。该性状是一个复杂性状,在个体水平上受多种遗传和环境因素的影响。 本研究之前的工作通过选择性发育起始时间差异较大的两种品系小鼠C3H/HeJ和C57BL/6作为亲本,产生F7代。通过STR, SNP位标利用特异性区段同类系的家系构建策略在小鼠X染色体上确定了一个和小鼠阴门开启相关的的Rs13483770～Rs29055848之间～1cM(物理距离1.7Mb)的区间长度QTL区段。 本研究为了更进一步精细定位小鼠X染色体上与性发育相关的候选基因,针对发育较早的C3H/HeJ品系小鼠,将该QTL内的12个候选基因(Mir505, Atp11c, Gm7073, Gm7076, Gm14661, Gm14662, Gm5637, Mcf2, F9, Fgf13, Gm715, Sox3)的编码序列及其5’上游5k列进行了序列测定。利用Lasergene7.0软件分析测序结果,首先通过SNP单倍型分析手段,初步排除了2个基因(Gm14662和F9)。后又进行反方向测序验证,排除了3个无差异基因(Fgfl3, Gm5637, Mcf2)。利用之前本实验室构建的特异性区段同类系小鼠进行目的片段的重测序分析,选出4个候选基因(Mir505, Atp11c, Gm7073, Gm14661)。最后,结合本实验室表型性状差异较大的4个野老鼠品系(CM25, JS11, JS13, PD5)在以上候选基因处测序,按表型差异特点进行序列差异分析。将另外2个候选基因(Gm7073, Gm14661)逐一排除。此时,该区段内与小鼠性发育启动时间相关的候选基因被锁定于Atp11c和Mir505。 本研究的结果初步确定了2个性发育启动基因,为今后其相关机制的研究打下了良好的基础。同时也有助于性发育启动相关通路调节的研究。所有这些工作均有助于帮助我们理解性发育复杂性状的整个生理过程和调控机制。
[Abstract]:Sexual development refers to the process of individual maturation and reproductive ability, which is influenced by many factors. The initiation time of sexual development is one of the most important traits in the process of sexual development. This trait is a complex trait which is influenced by many genetic and environmental factors at the individual level. In this study, two strains of mice, C3H/HeJ and C57BL/6, were used as parents to produce F7 generation. An interval length QTL region between Rs13483770~Rs29055848 and mouse vulvar opening-related Rs13483770~Rs29055848 (physical distance 1.7Mb) was determined on the X chromosome of mice by using the family construction strategy of the same family with specific regions by STR, SNP marker. In order to further map the candidate genes related to sexual development on the X chromosome of mice, 12 candidate genes (Mir505, Atp11c, Gm7073, Gm7076, Gm14661, Gm14662, Gm5637, Mcf2, F9, Mir505, Atp11c, Gm7073, Gm7076, Gm14661, Gm14662, Gm5637, Mcf2, F9, F9) in the QTL were selected for the early developing C3H/HeJ strain mice. The coding sequence of Fgf13, Gm715, Sox3) and its 5 'upstream 5k column were sequenced. Two genes (Gm14662 and F9) were initially excluded by SNP haplotype analysis using Lasergene7.0 software. Three indifference genes (Fgfl3, Gm5637, Mcf2) were excluded by reverse sequencing. Four candidate genes (Mir505, Atp11c, Gm7073, Gm14661) were selected by resequencing the target fragments from the mice of the same kind of specific region constructed in our laboratory. Finally, four strains of wild mice (CM25, JS11, JS13, PD5), which had great phenotypic differences in our laboratory, were sequenced at the above candidate genes, and the sequence differences were analyzed according to the phenotypic differences. The other two candidate genes (Gm7073, Gm14661) were excluded one by one. At this point, the candidate genes related to the initiation time of mouse sexual development in this region are locked to Atp11c and Mir505.. The results of this study provide a good basis for the further study of the related mechanism of personality development by identifying the promoter genes of personality development. At the same time, it is helpful to study the regulation of sexual development initiation-related pathway. All these work will help us to understand the whole physiological process and regulatory mechanism of complex traits of sexual development.
【学位授予单位】：东华大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R3416
本文编号：2458861
[Abstract]:Sexual development refers to the process of individual maturation and reproductive ability, which is influenced by many factors. The initiation time of sexual development is one of the most important traits in the process of sexual development. This trait is a complex trait which is influenced by many genetic and environmental factors at the individual level. In this study, two strains of mice, C3H/HeJ and C57BL/6, were used as parents to produce F7 generation. An interval length QTL region between Rs13483770~Rs29055848 and mouse vulvar opening-related Rs13483770~Rs29055848 (physical distance 1.7Mb) was determined on the X chromosome of mice by using the family construction strategy of the same family with specific regions by STR, SNP marker. In order to further map the candidate genes related to sexual development on the X chromosome of mice, 12 candidate genes (Mir505, Atp11c, Gm7073, Gm7076, Gm14661, Gm14662, Gm5637, Mcf2, F9, Mir505, Atp11c, Gm7073, Gm7076, Gm14661, Gm14662, Gm5637, Mcf2, F9, F9) in the QTL were selected for the early developing C3H/HeJ strain mice. The coding sequence of Fgf13, Gm715, Sox3) and its 5 'upstream 5k column were sequenced. Two genes (Gm14662 and F9) were initially excluded by SNP haplotype analysis using Lasergene7.0 software. Three indifference genes (Fgfl3, Gm5637, Mcf2) were excluded by reverse sequencing. Four candidate genes (Mir505, Atp11c, Gm7073, Gm14661) were selected by resequencing the target fragments from the mice of the same kind of specific region constructed in our laboratory. Finally, four strains of wild mice (CM25, JS11, JS13, PD5), which had great phenotypic differences in our laboratory, were sequenced at the above candidate genes, and the sequence differences were analyzed according to the phenotypic differences. The other two candidate genes (Gm7073, Gm14661) were excluded one by one. At this point, the candidate genes related to the initiation time of mouse sexual development in this region are locked to Atp11c and Mir505.. The results of this study provide a good basis for the further study of the related mechanism of personality development by identifying the promoter genes of personality development. At the same time, it is helpful to study the regulation of sexual development initiation-related pathway. All these work will help us to understand the whole physiological process and regulatory mechanism of complex traits of sexual development.
【学位授予单位】：东华大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R3416
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相关期刊论文 前2条
1 罗小平,王慕逖;性早熟诊断与治疗的现状和进展[J];临床儿科杂志;2002年09期
2 黄妤;黄国庆;;高分辨率熔解——SNP及突变研究的最新工具[J];生命的化学;2007年06期
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