Piedrahita J A, Weaks R, Petrescu A, Shrode T W, Derr J N, Womack J E
Department of Veterinary Anatomy and Public Health, College of Veterinary Medicine, Texas A&M University, College Station 77843-4458, USA.
Anim Genet. 1997 Feb;28(1):14-20. doi: 10.1111/j.1365-2052.1997.00054.x.
The bovine leukaemia inhibitory factor was isolated from a phage library and sequences for the gene, in addition to 1213 bp of 5' and 432 bp of 3' sequences, were obtained and compared with other mammalian leukaemia inhibitory factor genes. Comparisons indicated amino acid homologies ranging from 89.6% to 77.2% with the human and mouse homologues, respectively. Analysis of 500 bp of 5' regulatory regions indicated homologies ranging from 83.6% to 74.4% with the corresponding human and sheep sequences, respectively. Additionally, bovine leukaemia inhibitory factor-specific primers were prepared, and a panel of bovine x hamster somatic cell lines were analysed by the polymerase chain reaction (PCR). Data indicated 93% concordance of leukaemia inhibitory factor with aldehyde dehydrogenase 2 located on bovine chromosome 17, and concordance of 81% with myelin basic protein situated on bovine chromosome 24. Southern analysis of selected hybrids confirmed the PCR results, thus conclusively assigning the bovine leukaemia inhibitory factor gene to chromosome 17. Sequence analysis also revealed a microsatellite in intron 2 of the bovine leukaemia inhibitory factor. Analysis of this region by PCR in 22 unrelated Bos taurus and 19 unrelated Bos indicus cattle detected nine different alleles. Polymorphic information content values were 0.53 and 0.80 in B. taurus and B. indicus, respectively. Additionally, the same leukaemia inhibitory factor primers successfully detected allelic variants at this locus in Bos javanicus, Bos guarus and Bison bison but not in Odocoileus virginianus.
从噬菌体文库中分离出牛白血病抑制因子，并获得了该基因的序列，以及5'端1213 bp和3'端432 bp的序列，并与其他哺乳动物的白血病抑制因子基因进行了比较。比较结果表明，与人类和小鼠同源物的氨基酸同源性分别为89.6%至77.2%。对500 bp的5'调控区分析表明，与相应的人类和绵羊序列的同源性分别为83.6%至74.4%。此外，制备了牛白血病抑制因子特异性引物，并通过聚合酶链反应（PCR）分析了一组牛×仓鼠体细胞系。数据表明，白血病抑制因子与位于牛17号染色体上的醛脱氢酶2的一致性为93%，与位于牛24号染色体上的髓鞘碱性蛋白的一致性为81%。对选定杂种的Southern分析证实了PCR结果，从而最终将牛白血病抑制因子基因定位到17号染色体上。序列分析还揭示了牛白血病抑制因子内含子2中的一个微卫星。通过PCR对22头无关的黄牛和19头无关的瘤牛进行该区域分析，检测到9个不同的等位基因。黄牛和瘤牛的多态信息含量值分别为0.53和0.80。此外，相同的白血病抑制因子引物成功地在爪哇牛、大额牛和美洲野牛中检测到了该位点的等位基因变体，但在弗吉尼亚白尾鹿中未检测到。
