van Eijk M J, Russ I, Lewin H A
Department of Animal Sciences, University of Illinois, Urbana 61801.
Mamm Genome. 1993;4(2):113-8. doi: 10.1007/BF00290436.
The order and recombination fractions (theta) between the bovine major histocompatibility complex DRB3, DYA, and prolactin (PRL) genes were determined by typing of 254 sperm from a triply heterozygous bull. A recently developed method, primer extension preamplification (PEP), was used to amplify the bovine sperm genome prior to amplification of specific loci by the polymerase chain reaction (PCR). At least 28 copies of the DRB3, PRL, or DYA gene were obtained from 50 cycles of PEP. For sperm typing, alleles of each locus were discriminated by restriction endonuclease cleavage of PCR products and polyacrylamide gel electrophoresis of the restriction fragments. The most likely gene order is PRL-DRB3-DYA, with theta = 0.025 (+/- 0.012) and theta = 0.150 (+/- 0.024), respectively. The odds are 128:1 in favor of this order in comparison with the second most likely order DRB3-PRL-DYA. Our results demonstrate the power of sperm typing in concert with PEP for multilocus gene mapping.
通过对一头三重杂合公牛的254个精子进行分型，确定了牛主要组织相容性复合体DRB3、DYA和催乳素（PRL）基因之间的顺序和重组率（θ）。一种最近开发的方法，即引物延伸预扩增（PEP），在通过聚合酶链反应（PCR）扩增特定基因座之前用于扩增牛精子基因组。通过50个循环的PEP，至少获得了28个DRB3、PRL或DYA基因拷贝。对于精子分型，通过PCR产物的限制性内切酶切割和限制性片段的聚丙烯酰胺凝胶电泳来区分每个基因座的等位基因。最可能的基因顺序是PRL-DRB3-DYA，其θ值分别为0.025（±0.012）和0.150（±0.024）。与第二可能的顺序DRB3-PRL-DYA相比，支持该顺序的几率为128:1。我们的结果证明了精子分型与PEP协同用于多位点基因定位的能力。
