CHROMOSOME MAPPING IN DOMESTIC ANIMALS BY IN SITU HYBRIDIZATION

本研究旨在建立一种家畜染色体原位杂交定位基因的方法。^ 3h标记的LCAT（卵磷脂-胆固醇酰基转移酶）DNA，其位点已知（No. 3h）。16q22)，作为探针用于人染色体的原位杂交。然后，为了建立牛染色体原位杂交方法，以^ 3h标记的牛LDL（低密度脂蛋白）受体DNA作为探针。然而，由于该DNA可能不适合作为原位杂交探针，因此没有确定其位点。利用生物素标记的α -卫星DNA探针，利用荧光原位杂交技术（FISH）定位人类染色体上的基因。经异硫氰酸亲和素荧光素处理和碘化丙啶反染后，在染色体上检测到荧光信号。他们展示了α卫星DNA的位点。将建立的FISH方法应用于转基因大鼠染色体上的转基因定位。在YS新技术研究所将小鼠乳清酸性蛋白（m-WAP）插入原核大鼠卵中，获得转基因大鼠外周血淋巴细胞培养物的中期染色体。FISH前用tripsin技术观察到染色体呈g带状。转基因（m-WAP）定位于no. q14位点。6染色体核型分析。认为本研究对家畜基因定位有一定的贡献。

The present study aimed to establish a method of localization of genes in chromosomes of domestic animals by in situ hybridization.Preliminarily, ^3H-labeled LCAT (lecithin-cholesterol acyltransferase)DNA,whose locus has been known(No.16q22), was used for in situ hybridization of human chromosomes as a probe. Then, in order to establish a method of in situ hybridization in bovine chromosomes, ^3H-labeled bovine LDL (low-density lipoprotein)receptor DNA was used as a probe. However, the locus was not determined because this DNA might be not suitable for in situ hybridization as the probe.Fluorescence in site hybridization(FISH) was attempted to localize a gene on human chromosomes using alpha-satellite DNA probe labeled with biotin. After treatment with avidin-fluorescein isothiocyanate and counterstain with propidium iodide, fluorescent signals were detected on chromosomes. They have shown the loci of alpha-satellite DNA.The established FISH method was applied to localization of a transgene on chromosomes of transgenic rats. Metaphase chromosome spreads were obtained from peripheral blood lymphocyte cultures of transgenic rats, which were produced by insertion of mouse whey acidic protein(m-WAP) into pronuclear rat eggs in YS New Techonology Institute. Chromosomes were G-banded by tripsin technique before FISH.The transgene (m-WAP)was localized at q14 site of no.6 chromosome by karyotype analysis.It is considered that the present study contributes to the gene mapping in domestic animals.