中国人群中STK15基因内两种非同义SNP的连锁不平衡和单体型分析(英文)  

Linkage Disequilibrium and Haplotype Analysis of Two Single Nucleotide Polymorphisms in STK15 in Chinese

作　　者：陈立[1] 敖雪[1] 任群[1] 王振宁[1] 鲁翀[1] 徐岩[1] 姜莉[1] 罗阳[1] 徐惠绵[1] 张学[1,2] 

机构地区：[1]中国医科大学医学基因组学研究室卫生部细胞生物学重点实验室 [2]中国医学科学院中国协和医科大学基础医学研究所医学遗传学系医学分子生物学国家重点实验室,北京100005

出　　处：《Acta Genetica Sinica》2005年第4期331-336,共6页

基　　金：国家杰出青年科学基金 (编号: 30125017 )30125017 ) ]~~

摘　　要：STK15基因编码一种丝氨酸苏氨酸蛋白激酶,哺乳动物细胞中其过量表达将导致中心体扩增、染色体不稳定和细胞癌变。STK15基因外显子 3中有 3种非同义单核苷酸多态 (SNP),即: 91A→T(I31F)、169G→A(V57I)和 311C→T(S104L)。新近研究发现, 91A→T与人类肿瘤遗传易感性相关。应用PCR RFLP技术确定了 91A→T(I31F)和 169G→A(V57I)两种SNP在中国人群中的基因型和单体型。采用巢式PCR方法扩增了193例正常个体的DNA样品,通过错配正向巢式内引物引入EcoRⅠ酶切位点。巢式PCR扩增产物用限制性内切酶EcoRⅠ和AccⅡ双酶切消化,其中EcoRⅠ能酶切 91A,AccⅡ能切开 169G,用聚丙烯酰胺凝胶电泳银染法鉴定双酶切结果,发现了 4种可能的单体型中的 3种,其单体型频率分别为:p( 91A 169G) =68 65%,p( 91T 169A) = 10 88%,p(91T 169G) =20 47%,p(91A 169A) =0%;它们组成的 6种基因型及频率分别为: 91A 169G/91A 169G ( 46 11% ), 91A 169G/91T 169A( 14 51% ), 91A 169G/91T 169G( 30 57% ), 91T 169G/91T 169G(3 11% ), 91T 169G/91T 169A( 4 15% ), 91T 169A/91T 169A( 1 55% )。等位基因及单体型数据分析结果表明, 91A→T(I31F)和 169G→A(V57I)之间存在连锁不平衡。STK15 (Serine/Threonine protein kinase 15) is a serine/threonine kinase encoding gene,whose overexpression in mammalian cells leads to centrosome amplification,chromosomal instability,and oncogenic transformation.91A→T,a single nucleotide polymorphism (SNP) in exon 3 of STK15,has recently been shown to be associated with human cancer susceptibility.Within exon 3 of STK15,there are three nonsynonymous SNPs: 91A→T,169G→A and 311C→T.We have determined STK15 genotypes and haplotypes composed of 91A→T and 169G→A by PCR-RFLP in a randomly sampled cohort of 193 normal individuals from Northeast China.DNA samples from all individuals were subjected to first round of PCR using a pair of specific primers.For the subsequent nested PCR,a mismatch forward primer,which could introduce an EcoRⅠ restriction site to the 91A allele,was included.The nested PCR products were digested with the restriction endonucleases EcoRⅠ and AccⅡ.The double restriction digests were separated by polyacrylamide gel electrophoresis.Three haplotypes,91A-169G,91T-169A and 91T-169G,were detected and their frequencies were 68.65%,10.88% and 20.47%,respectively.Six genotypes composed of the above three haplotypes were found,and their frequencies were 91A-169G/91A-169G (46.11%),91A-169G/91T-169A (14.51%),91A-169G/91T-169G (30.57%),91T-169G/91T-169G (3.11%),91T-169G/91T-169A (4.15%), 91T-169A/91T-169A (1.55%).Whereas no 91A-169A haplotype was detected in all individuals examined in the current study,linkage disequilibrium (LD) between the two SNPs was found.

关 键 词：STK15基因 单核苷酸多态 单体型 连锁不平衡 

分 类 号：Q987[生物学—遗传学]