A PstI DNA polymorphism in the human stefin A gene (STF 1).
A PstI DNA polymorphism in the human stefin A gene (STF 1).
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人类 Stefin A 基因 (STF 1) 中的 PstI DNA 多态性。
DOI：
10.1093/nar/19.7.1722-a
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发表时间：
1991
影响因子：
14.9
通讯作者：
Fong,D
中科院分区：
文献类型：
--
作者：
Hsieh,WT;Barrick,JL;Berrettini,WH;Chan,MM;Fong,D
PCR引物：2E12B#1：5 '-TGGAACACTCAGGCGA-3' 2E12B#2：5 '-CCAGAGCCACTTTCTAC-3'多态性：在DNA测序凝胶上分辨出10个等位基因片段。等位基因片段（nt）的长度为：A1 = 193，A2= 191，A3= 189，A4= 187，A5= 185，A6= 183，A7= 181，A8= 161，A9= 159，A10 = 155。频率：52名无关白种人的等位基因频率为：Al=。01，A2=. 01，A3=. 11，A4=. 13，A5=. 26，A6=. 02，A7=. 06，A8=. 01，A9=. 38，AIO=. 01; PIC= 0.72。染色体定位和孟德尔遗传：来自两个体细胞杂种组的基因组DNA的PCR表明定位于14号染色体。荧光原位杂交显示定位于14 q24。3.在8个CEPH家族中对基因座D14 S24（探针CRI-C70）进行的具有85个信息性减数分裂的连锁分析在0= 0.3时给出了2.95的最大LOD得分。在所有病例中均观察到孟德尔遗传。PCR条件：PCR反应在总体积为25 μ l的容器中进行，该容器含有：25 ng基因组DNA，10 pmole的每种引物，1.5 mM MgCl 2，200 sM dNTPs，50 mM KCl，10 mM Tris-Cl，pH 8.3，0.6单位Taq聚合酶（Ampli-Taq）和0.01%明胶。扩增进行35个循环，94 ℃变性60秒，54 ℃退火60秒，72 ℃延伸30秒。该标记位点可作为14号染色体遗传连锁图的一个有用的指示标记。鸣谢：这项工作得到了NIH Grant HG 00022的支持。夏尔马是俄勒冈州医学研究基金会奖学金的获得者。我们感谢H. Lehrach提供粘粒ICRFclO 2 E1222。参考文献：1）Roberts，L.（1990）Science 248，805.
PCR Primers: 2E12B# 1: 5'-TGGAACACTCAGGCGA-3'2E12B# 2: 5'-CCAGAGCCACTTTCTAC-3'Polymorphism: Ten allelicfragments were resolved on DNA sequencing gels. Lengths of allelic fragments (nt) were: Al= 193, A2= 191, A3= 189, A4= 187, A5= 185, A6= 183, A7= 181, A8= 161, A9= 159, AlO= 155. Frequencies: Allele frequencies in 52 unrelated Caucasians were: Al=. 01, A2=. 01, A3=. 11, A4=. 13, A5=. 26, A6=. 02, A7=. 06, A8=. 01, A9=. 38, AIO=. 01; PIC= 0.72.Chromosomal Localization and Mendelian Inheritance: PCR of genomic DNAs from two somatic cell hybrid panels indicated localization to chromosome 14. Fluorescent in situ hybridization indicated localization to 14q24. 3. Linkage analysis in 8 CEPH families with 85 informative meioses against locus D14S24 (probe CRI-C70) gave a maximum LOD score of 2.95 at 0= 0.3. Mendelian inheritance was observed in all cases. PCR Conditions: PCR reactions are carried out in a total volume of 25 A1 containing: 25 ng genomic DNA, 10 pmole of each primer, 1.5 mM MgCl2, 200 sM dNTPs, 50 mM KCl, 10 mM Tris-Cl, pH 8.3, 0.6 units Taq polymerase (Ampli-Taq) and 0.01% gelatin. Amplification is for 35 cycles with denaturation at 94 C for 60 seconds, annealing at 54 C for 60 seconds and extension at 72 C for 30 seconds. Comnments: This marker locus will be a useful index marker (1) for the genetic linkage map of chromosome 14. Acknowledgements: Thiswork was supported by NIH Grant HG00022. V. Sharma is the recipient of a fellowship from the Medical Research Foundation of Oregon. We thank H. Lehrach for providing cosmid ICRFclO2E1222. Reference: 1) Roberts, L.(1990) Science 248, 805.