实验用狨猴微卫星引物筛选及遗传多样性分析
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摘要
目的筛选、优化狨猴微卫星DNA引物,用以对中国医学科学院医学实验动物研究所引进的实验用狨猴种群进行遗传学分析及评估。方法用筛选的20对狨猴微卫星引物,对随机抽取的30只狨猴血液样本进行基因组DNA提取及PCR扩增,扩增产物经电泳鉴定后进行STR扫描检测,用Popgene1.32软件对STR扫描结果进行数据处理和分析。结果 20对微卫星引物均呈现出遗传多样性,共检测147个等位基因,观察等位基因数5~10个,平均7.35个;有效等位基因数2.2500~6.3830个,平均4.0402个;观察杂合度0.000~0.4667,平均0.1533;期望杂合度0.1424~0.4350,平均0.2506;香隆指数1.2242~2.0324,平均1.5949;多态信息含量0.5366~0.8254,平均0.7053。结论本文筛选的20对狨猴微卫星引物具有高度遗传多样性,均处于Hardy-Weinberg平衡状态。
Objective To screen and optimize the microsatellite DNA primers of the laboratory common marmoset,analyze and evaluate the population genetic quality for the marmosets(Callithrix jacchus) introduced into the Institute of Medical Laboratory Animal science,Chinese Academy of Medical Sciences. Methods A total of 30 marmosets were randomly chosen, and their genome DNA from blood was extracted using phenol/chloroform method. The microsatellite DNA was amplified using standard polymerase chain reaction(PCR). The amplification products were tested by STR scanning after 2% agrose gel and 8% PAGE electrophoresis. The data processing and genetic analysis were completed using the Popgene1. 32 software. Results A total of 20 pairs of microsatellite loci showed genetic polymorphism,and 147 alleles were detected. The number of allele was 5 to 10,average 7. 35. The effective allele was2. 2500 to 6. 3830,average 4. 0402. The observed heterozygosity was 0. 000 to 0. 4667,average 0. 1533. The expected heterozygosity was 0. 1424 to 0. 4350,average 0. 2506. The Shannon diversity index was 1. 2242 to 2. 0324,average1. 5949. The polymorphic information content was 0. 5366 to 0. 8254,average 0. 7053. Conclusions The 20 pairs of marmoset microsatellite primers are genetically highly diverse and are in a Hardy-Weinberg equilibrium.
Objective To screen and optimize the microsatellite DNA primers of the laboratory common marmoset,analyze and evaluate the population genetic quality for the marmosets(Callithrix jacchus) introduced into the Institute of Medical Laboratory Animal science,Chinese Academy of Medical Sciences. Methods A total of 30 marmosets were randomly chosen, and their genome DNA from blood was extracted using phenol/chloroform method. The microsatellite DNA was amplified using standard polymerase chain reaction(PCR). The amplification products were tested by STR scanning after 2% agrose gel and 8% PAGE electrophoresis. The data processing and genetic analysis were completed using the Popgene1. 32 software. Results A total of 20 pairs of microsatellite loci showed genetic polymorphism,and 147 alleles were detected. The number of allele was 5 to 10,average 7. 35. The effective allele was2. 2500 to 6. 3830,average 4. 0402. The observed heterozygosity was 0. 000 to 0. 4667,average 0. 1533. The expected heterozygosity was 0. 1424 to 0. 4350,average 0. 2506. The Shannon diversity index was 1. 2242 to 2. 0324,average1. 5949. The polymorphic information content was 0. 5366 to 0. 8254,average 0. 7053. Conclusions The 20 pairs of marmoset microsatellite primers are genetically highly diverse and are in a Hardy-Weinberg equilibrium.
引文
[1]田岛嘉雄.实验动物的生物学特性资料[M].第一版.实验动物中央研究所,1989:486-488.
[2]Kishi N,Sato K,Sasaki E,et al.Common marmoset as a new model animal for neuroscience research and genome editing technology[J].Dev Growth Differ,2014,56(1):53-62.
[3]Mansfield K.Marmoset models commonly used in biomedical research[J].Comp Med,2003,53(4):383-392.
[4]马波,胡彩珍,何丽芳,等.精制甲型肝炎灭活疫苗(YN5株)狨猴试验安全性和免疫原性[J].中国生物制品学杂志,2004,17(1):36-38.
[5]李婷婷,帅丽芳,陈姿喧,等.丙型肝炎病毒结构蛋白的嵌合病毒狨猴感染模型[J].中国输血杂志,2010,23(s1):167.
[6]殷森.普通棉耳绒猴MHC-I型基因多态性特征分析[D].南方医科大学,2013.
[7]徐玉霞.普通棉耳狨猴MHC-I限制性等位基因抗原分子特征分析[D].南方医科大学,2015.
[8]石亮,张晨,向志光,等.腺相关病毒介导的狨猴P53基因沉默[J].中国比较医学杂志,2016,26(4):53-57.
[9]邓怡晨,张晨,向志光,等.狨猴B2m基因沉默位点在细胞水平的验证[J].中国比较医学杂志,2017,27(5):37-41.
[10]王越甲,杨炜峰,车路平,等.利用优选的微卫星标记建立常用大小鼠品系的遗传监测体系[J].实验动物科学,2014,31(3):5-9,14.
[11]商海涛,魏泓,岳秉飞,等.应用微卫星标记对三个昆明小鼠封闭群的遗传学研究[J].实验动物科学,2009,26(2):1-6.
[12]商海涛,魏泓,岳秉飞,等.应用微卫星DNA标记对Wistar和SD大鼠封闭群的遗传学研究[J].分子细胞生物学报,2008,41(1):28-34.
[13]丁贤明,钱宝珍,Junichiro M,等.长爪沙鼠的遗传多样性分析[J].遗传,2008,30(7):877-884.
[14]魏杰,王吉星,王洪,等.房山封闭群小型猪微卫星位点的测定[J].中国比较医学杂志,2013,23(11):7-12.
[15]禹文海,和占龙,鲁绍雄,等.基于微卫星DNA标记的恒河猴遗传多样性研究[J].中国比较医学杂志,2013,23(3):21
[16]李瑞生,李晓娟,高蓉,等.食蟹猴微卫星DNA标记遗传监测及多态性分析[J].中国比较医学杂志,2011,21(8):27-30.
[17]Raveendran M,Tardif S,Ross CN,et al.Polymorphic microsatellite loci for the common marmoset(Callithrix jacchus)designed using a cost-and time-efficient method[J].Am J Primatol,2008,70(9):906-910.
[18]Katoh H,Takabayashi S,Itoh T.Development of microsatellite DNA markers and their chromosome assignment in the common marmoset[J].Am J Primatol,2009,71(11):912-918.
[19]左宝芬,杜小燕,霍学云,等.五个C57BL/6J小鼠生产群的微卫星检测及遗传学质量分析[J].实验动物与比较医学,2012,32(1):51-55.
[20]王洪,杜小燕,徐平,等.上海KM小鼠种子群体遗传状况分析[J].中国比较医学杂志,2014,24(12):27-32.
[21]Botstein D,White RL,Skolnick M,et al.Construction of a genetic linkage map in man using restriction fragment length polymorphisms[J].Am J Hum Genet,1980,32(3):314-331.
[2]Kishi N,Sato K,Sasaki E,et al.Common marmoset as a new model animal for neuroscience research and genome editing technology[J].Dev Growth Differ,2014,56(1):53-62.
[3]Mansfield K.Marmoset models commonly used in biomedical research[J].Comp Med,2003,53(4):383-392.
[4]马波,胡彩珍,何丽芳,等.精制甲型肝炎灭活疫苗(YN5株)狨猴试验安全性和免疫原性[J].中国生物制品学杂志,2004,17(1):36-38.
[5]李婷婷,帅丽芳,陈姿喧,等.丙型肝炎病毒结构蛋白的嵌合病毒狨猴感染模型[J].中国输血杂志,2010,23(s1):167.
[6]殷森.普通棉耳绒猴MHC-I型基因多态性特征分析[D].南方医科大学,2013.
[7]徐玉霞.普通棉耳狨猴MHC-I限制性等位基因抗原分子特征分析[D].南方医科大学,2015.
[8]石亮,张晨,向志光,等.腺相关病毒介导的狨猴P53基因沉默[J].中国比较医学杂志,2016,26(4):53-57.
[9]邓怡晨,张晨,向志光,等.狨猴B2m基因沉默位点在细胞水平的验证[J].中国比较医学杂志,2017,27(5):37-41.
[10]王越甲,杨炜峰,车路平,等.利用优选的微卫星标记建立常用大小鼠品系的遗传监测体系[J].实验动物科学,2014,31(3):5-9,14.
[11]商海涛,魏泓,岳秉飞,等.应用微卫星标记对三个昆明小鼠封闭群的遗传学研究[J].实验动物科学,2009,26(2):1-6.
[12]商海涛,魏泓,岳秉飞,等.应用微卫星DNA标记对Wistar和SD大鼠封闭群的遗传学研究[J].分子细胞生物学报,2008,41(1):28-34.
[13]丁贤明,钱宝珍,Junichiro M,等.长爪沙鼠的遗传多样性分析[J].遗传,2008,30(7):877-884.
[14]魏杰,王吉星,王洪,等.房山封闭群小型猪微卫星位点的测定[J].中国比较医学杂志,2013,23(11):7-12.
[15]禹文海,和占龙,鲁绍雄,等.基于微卫星DNA标记的恒河猴遗传多样性研究[J].中国比较医学杂志,2013,23(3):21
[16]李瑞生,李晓娟,高蓉,等.食蟹猴微卫星DNA标记遗传监测及多态性分析[J].中国比较医学杂志,2011,21(8):27-30.
[17]Raveendran M,Tardif S,Ross CN,et al.Polymorphic microsatellite loci for the common marmoset(Callithrix jacchus)designed using a cost-and time-efficient method[J].Am J Primatol,2008,70(9):906-910.
[18]Katoh H,Takabayashi S,Itoh T.Development of microsatellite DNA markers and their chromosome assignment in the common marmoset[J].Am J Primatol,2009,71(11):912-918.
[19]左宝芬,杜小燕,霍学云,等.五个C57BL/6J小鼠生产群的微卫星检测及遗传学质量分析[J].实验动物与比较医学,2012,32(1):51-55.
[20]王洪,杜小燕,徐平,等.上海KM小鼠种子群体遗传状况分析[J].中国比较医学杂志,2014,24(12):27-32.
[21]Botstein D,White RL,Skolnick M,et al.Construction of a genetic linkage map in man using restriction fragment length polymorphisms[J].Am J Hum Genet,1980,32(3):314-331.