遗传学报
遺傳學報
유전학보
ACTA GENETICA SINICA
2005年
4期
331-336
,共6页
陈立%敖雪%任群%王振宁%鲁翀%徐岩%姜莉%罗阳%徐惠绵%张学
陳立%敖雪%任群%王振寧%魯翀%徐巖%薑莉%囉暘%徐惠綿%張學
진립%오설%임군%왕진저%로충%서암%강리%라양%서혜면%장학
STK15基因%单核苷酸多态%单体型%连锁不平衡
STK15基因%單覈苷痠多態%單體型%連鎖不平衡
STK15기인%단핵감산다태%단체형%련쇄불평형
STK15%SNP%haplotype%linkage disequilibrium
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基因編碼一種絲氨痠囌氨痠蛋白激酶,哺乳動物細胞中其過量錶達將導緻中心體擴增、染色體不穩定和細胞癌變.STK15基因外顯子3中有3種非同義單覈苷痠多態(SNP),即:91A→T(I31F)、169G→A(V57I) 和311C→T(S104L).新近研究髮現,91A→T與人類腫瘤遺傳易感性相關.應用PCR-RFLP技術確定瞭91A→T(I31F)和 169G→A(V57I)兩種SNP在中國人群中的基因型和單體型.採用巢式PCR方法擴增瞭193例正常箇體的DNA樣品,通過錯配正嚮巢式內引物引入EcoRⅠ酶切位點.巢式PCR擴增產物用限製性內切酶EcoRⅠ和AccⅡ雙酶切消化,其中EcoRⅠ能酶切91A,AccⅡ能切開169G,用聚丙烯酰胺凝膠電泳銀染法鑒定雙酶切結果,髮現瞭4種可能的單體型中的3種,其單體型頻率分彆為:p(91A-169G)=68.65%,p(91T-169A)= 10.88%,p(91T-169G)=20.47%,p(91A-169A)=0%;它們組成的6種基因型及頻率分彆為: 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%).等位基因及單體型數據分析結果錶明,91A→T(I31F)和 169G→A(V57I)之間存在連鎖不平衡.
STK15기인편마일충사안산소안산단백격매,포유동물세포중기과량표체장도치중심체확증、염색체불은정화세포암변.STK15기인외현자3중유3충비동의단핵감산다태(SNP),즉:91A→T(I31F)、169G→A(V57I) 화311C→T(S104L).신근연구발현,91A→T여인류종류유전역감성상관.응용PCR-RFLP기술학정료91A→T(I31F)화 169G→A(V57I)량충SNP재중국인군중적기인형화단체형.채용소식PCR방법확증료193례정상개체적DNA양품,통과착배정향소식내인물인입EcoRⅠ매절위점.소식PCR확증산물용한제성내절매EcoRⅠ화AccⅡ쌍매절소화,기중EcoRⅠ능매절91A,AccⅡ능절개169G,용취병희선알응효전영은염법감정쌍매절결과,발현료4충가능적단체형중적3충,기단체형빈솔분별위:p(91A-169G)=68.65%,p(91T-169A)= 10.88%,p(91T-169G)=20.47%,p(91A-169A)=0%;타문조성적6충기인형급빈솔분별위: 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%).등위기인급단체형수거분석결과표명,91A→T(I31F)화 169G→A(V57I)지간존재련쇄불평형.
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.
