Tetranucleotide repeat polymorphism at the human thyroid peroxidase (hTPO) locus.
Tetranucleotide repeat polymorphism at the human thyroid peroxidase (hTPO) locus.
复制标题
人甲状腺过氧化物酶 (hTPO) 位点的四核苷酸重复多态性。
DOI：
10.1093/hmg/1.2.137
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发表时间：
1992
影响因子：
3.5
通讯作者：
Donis-Keller,H
中科院分区：
文献类型：
--
作者：
Anker,R;Steinbrueck,T;Donis-Keller,H
染色体定位：通过体细胞杂交的Southern blot杂交，hTPO基因被定位在2p上（1,2）。我们通过与D2S1的遗传连锁进一步将hTPO定位为2p23-2pter，之前通过原位杂交定位为2p23-2pter(3)。图2显示了用CRI-MAP程序包构建的多点遗传连锁图谱(4)、来自8个CEPH家系的sRA-1 （TPO）基因型和来自CEPH数据库的另外3个标记的基因型数据（release 5）。其他评论：PCR反应在10毫升的总体积中进行，其中含有PCR缓冲液（10 mM Tris-HCl pH: 8.3, 50 mM KC1, 1.5 mM MgCl2）， 100 ng基因组DNA，每个未标记引物1 tiM， 800 jtM dNTPs， 0.5 U Taq聚合酶（Boehringer Mannheim GmbH）。AATG链引物5′32P端标记，以0.1 jtM的浓度加入到反应混合物中。反应混合物在94℃下变性1分钟，63℃下退火30秒，72℃下延长90秒，共培养22个循环。
Chromosomal Localization: The hTPO gene has been localized to 2p by Southern blot hybridization of somatic cell hybrids (1, 2). We have further subregionally localized hTPO to 2p23-2pter by genetic linkage to D2S1, previously mapped to 2p23-2pter by in situ hybridization (3). Figure 2 shows a multipoint genetic linkage map constructed with the CRI-MAP program package (4), genotypes from 8 CEPH pedigrees for sRA-1 (TPO) and genotype data for 3 additional markers from the CEPH database (release 5).Other Comments: PCR reactions were performed in a total volume of 10 jil containing PCR buffer (10 mM Tris-HCl pH: 8.3, 50 mM KC1, 1.5 mM MgCl2), 100 ng genomic DNA, 1 tiM each unlabeled primer, 800 jtM dNTPs, 0.5 U Taq polymerase (Boehringer Mannheim GmbH). In addition, the AATG strand primer was 5'32P end-labelled and added to the reaction mixture at a concentration of 0.1 jtM. The reaction mixture was incubated for 22 cycles with denaturation at 94 C for 1 min, annealing at 63 C for 30 sec, and extension at 72 C for 90 sec.