【摘要】 为了建立鲤IGF2b （insulin-like growth factor 2）基因全基因组信息SNPs （single nucleotide polymorphism）的获取体系,并验证该获取体系在黄河鲤新品系选育中运用的适用性,本研究首先对10个不同鲤品种的33个鲤个体重测序数据进行分析,整体上了解IGF2b基因的单核苷酸位点及其在基因组上的分布情况。然后在基因组DNA水平上分段获取IGF2b基因的SNPs信息,并以不同的鲤品种验证引物的适用性,最后在黄河鲤新品系中检测其应用。研究结果:获得8对聚合酶链式反应（polymerase chain reaction, PCR）引物进行分段克隆的结果,并通过直接测序和限制性片段长度多态性来获取相应的SNPs。经过直接测序,本文对黄河鲤、建鲤、黄河鲤和建鲤的正反交后代进行PCR检测发现条带单一,目地片段正确;结合黄河鲤新品系的体重数据,本文又以IGF2b^3#引物检测到一个与体重明显相关的SNPs,同时检测到一个与该基因表达量相关的另一个SNPs。本研究的方法能够在该基因全基因组范围内开展鲤分子育种中SNPs的检测,并验证发现黄河鲤新品系IGF2b^3#引物227号位点处,出现的突变位点使体重降低,且有一处SNPs与其表达量有关。这将为其他物种、其他功能基因的多态性分子标记研究提供思路。更多还原

【Abstract】 In order to establish an SNP（single nucleotide polymorphism） acquisition system for the IGF2b （insulin-like growth factor 2） gene, a total of 33 DNA samples extracted from 10 different carp species were sequenced and the data were analyzed to verify the function and efficacy of the breeding process for the new Huanghe carp strain. By determining the position and distribution of SNPs on the genome and obtaining the IGF2b gene by segmentation at the genomic DNA level in this way, the applicability of primers from different carp species for use in Huanghe carp new strains was verified. As a result of the segmentation at the genomic DNA level, eight pairs of polymerase chain reaction（PCR） primers were obtained, and the corresponding SNPs were determined by direct sequencing and restriction fragment length polymorphisms. Furthermore, direct sequencing showed a single band from the PCR of Huanghe carp and Jian carp as well as the progenies resulting from their reciprocal crosses, which allowed us to confirm the validity of the target fragment. Finally, analysis of the Huanghe carp body weight data combined with the latest results allowed the detection of an SNP significantly associated with the body weight using IGF2b^3# primers. Likewise, another SNP was detected in association with the expression level of this gene.The methods used in this study were able to detect the SNPs of genes in the molecular breeding of carp within the range of the whole genome, to verify that the mutation at the No. 227 site on the IGF2b^3# primer of Huanghe carp led to reduced body weight, and to identify another SNP related to the expression level of IGF2b. This will provide the background for the study of polymorphic molecular markers and other functional genes in other species.更多还原