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野猪、家猪及其杂种猪产仔数相关基因的比较研究
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摘要
母猪的产仔数性状是一个遗传力很低的数量性状(h2=0.1),用传统的选育方法提
    高母猪产仔数收效甚微,从二十世纪八十年代起,以 Rothschild 为代表的一批科学家,
    通过采用遗传标记和标记辅助选择的方法,已经陆续发现了多个与产仔数性状有关的候
    选基因。
     因野猪、家猪及其杂种猪在产仔数性状上差异极其显著,因此,本实验通过对其 5
    个与产仔数性状相关的候选基因—ESR、FSHβ、LH、PRLR 和 RYR1,及其碱基序列
    的差异比较,比较因其碱基差异所造成的产仔数差异的 DNA 分子机制。家猪以地方品
    种东北民猪、引进品种长白猪和培育品种双肌臀大白猪为代表,主要采用了 PCR 和
    PCR-SSCP 技术。
     结果表明,基因克隆的部分序列中,三者在碱基序列上并不存在显著差异,仅存在
    个别碱基的差异,但由于有些差异碱基位于外显子处,因此不能确定这种碱基的变化是
    否会导致关键氨基酸的改变,造成功能上的变化,因此对存在变化的位点,在其两端设
    计引物,进行 SSCP 检测,来检验这种变化是否在每个猪种内都存在。结果:ESR第二
    对引物扩增的片段存在一个点突变 A→G,导致氨基酸 Arg→His 的变化,在家猪和野
    猪中均不存在 BB 型个体,但野猪中 B 基因的频率明显高于家猪的;FSHβ第一对引物
    扩增的片段存在两个点突变 C→T,导致氨基酸 Phe→Ser 的变化,G→A 的突变导致氨
    基酸 Ala→Thr 的变化;基因型检测结果,除双肌臀不存在 AA 型个体外,三种基因型
    在各个猪种内都有分布,不存在差异;FSHβ第三对引物扩增的片段存在一个点突变 C
    →T,无氨基酸变化,因此认为不存在差异;PRLR 第二对引物扩增的片段存在两个点
    突变 C→T 无氨基酸的变化,T→C 导致氨基酸 Val→Ala 的变化,统计分析的结果是杂
    种猪中没有 BB、CC 型个体,野猪中没有 BB 型,家猪中虽然都有,但是 BB、CC 型
    的个体也很少,但总的看来,这三者在 PRLR 基因的该片段上还是存在一定差异的;
    LH 第三对引物扩增的片段存在两个点突变,T→G、C→T,无氨基酸的变化,不存在
    差异;RYR1 第三对引物所扩增的片段存在一个点突变 C→T,无氨基酸的改变,三者
    不存在差异。综合起来考虑,野猪、家猪及其杂种猪在本实验所做的这五个基因的部分
    序列上存在一定的差异。
The litter size of pig is a low heritance quantitative trait(h2=0.1),in conventional method
    to improve this trait almost have no effect.Since the 1980’s, a set of scientist as well as
    Rothschild act as deputation,through the method of genetic marker and marker assited
    selection,had found some candidate genes about litter size trait.
     Because there is a most different between wild boar and pig in this trait,we wanted to
    through comparing the base sequence of wild boar and pig in these gene which had been
    considered as the candidate gene-ESR、FSH?、LH、PRLR and RYR1.The deputation of pig
    is native pig breeds of Min Pig,introduced pig breeds of Landrace and developed pig breeds
    of double muscle pig.Using technology of PCR-SSCP.
     Result:there is not most different in base sequence in large PCR fragment,only have
    some base different.But some different bases are locus on exon,they may lead to some crucial
    amino acid change and function change.We design primer in changed points to examine
    whether these change are all in three pig, PCR-SSCP technology was applied to test.Result:
    There is one mutant site in the fragment of the second primer of ESR: A→G,it lead amino
    acid change: Arg→His,both pig and wild boar are all have no BB genotype individual,but the
    frequence of B gene of wild boar is more higher than pig’s.There is two mutant site in the
    fragment of the first primer of FSHβ: C→T, G→A,it lead to amino acid change: Phe→Ser,
    Ala→Thr, they all have three genotype except shuangjitun has no AA.There is one mutant site
    in the fragment of the third primer of FSHβ:C→T,no amino acid change,so I think there is
    no difference in three pig breeds.There is two mutant site in the fragment of the third primer
    of LH: T→G,C→T, no amino acid change,no difference.There is two mutant site in the
    fragment of the second primer of PRLR: C→T,no amino acid change, T→C it lead to amino
    acid change: Val→Ala,the result of statistical analysis is hybrid pig have no BB,CC genotype
    individual,wild boar have no BB genotype,pig all have,but the individual of BB,CC are
    littile,too.Taking one with another,it is different in this fragment of PRLR gene among these
    pig breeds.There is one mutant site in the fragment of the third primer of RYR1: C→T, no
    amino acid change,no difference.Take into account whole,there is some difference in the five
    gene of part sequence among wild boar,pig and hybrid pig.
     II
    
    
    ABSTRACT
     Master candidate:Zhang Dongjie
     Specialty:Animal Genetics and Breeding
     Supervisor:Liu Di
引文
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