Takao S, Mykytyn K, Jacob C O
Institute of Immunology and Biological Sciences, Syntex Research, Palo Alto, CA 94303.
Immunogenetics. 1993;37(3):199-203. doi: 10.1007/BF00191885.
The introduction of the polymerase chain reaction (PCR) provides an entirely new means of analyzing DNA polymorphism and makes practical the analysis of length variation in simple-sequence tandem repeats of dinucleotides. In the process of cloning and sequencing the mouse genomic DNA for tumor necrosis factor (TNF) receptors type 1 and type 2, we identified two simple dinucleotide repeats within the noncoding regions of TNF receptor type 1 and three such sequences within TNF receptor type 2. PCR analysis of these sequences, using genomic DNA from 21 different inbred and wild mouse strains, as demonstrated by running the amplified products on sequencing gels, showed that the repeats are highly polymorphic. We identified seven alleles of TNF receptor type 2 and five alleles of TNF receptor type 1. Using these polymorphic markers in two sets of recombinant inbred strains of mice, the chromosomal localization of Tnfr-1 was mapped to mouse chromosome 6 and Tnfr-2 was located to the distal portion of mouse chromosome 4.
聚合酶链反应(PCR)的引入为分析DNA多态性提供了一种全新的方法,并使对二核苷酸简单序列串联重复中的长度变异分析变得切实可行。在克隆和测序小鼠1型和2型肿瘤坏死因子(TNF)受体的基因组DNA的过程中,我们在1型TNF受体的非编码区鉴定出两个简单的二核苷酸重复序列,在2型TNF受体中鉴定出三个这样的序列。使用来自21种不同近交系和野生小鼠品系的基因组DNA对这些序列进行PCR分析,通过在测序凝胶上运行扩增产物表明,这些重复序列具有高度多态性。我们鉴定出2型TNF受体的七个等位基因和1型TNF受体的五个等位基因。利用这些多态性标记物在两组重组近交系小鼠中,将Tnfr - 1的染色体定位到小鼠6号染色体,将Tnfr - 2定位到小鼠4号染色体的远端部分。