Chowdhary B P, Thomsen P D, Harbitz I, Landset M, Gustavsson I
Department of Animal Breeding and Genetics, Swedish Agricultural University, Uppsala.
Cytogenet Cell Genet. 1994;67(3):211-4. doi: 10.1159/000133825.
Fluorescence in situ hybridization (FISH) was applied, using genomic DNA clones, to precisely localize the genes for GPI, CRC, LIPE, and GH on pig chromosomes. The porcine CRC gene was localized to band 6q12 using both genomic and cDNA clones. The GPI and LIPE genes, which are closely linked to the CRC gene, were also mapped to the same band (6q12), using genomic lambda clones. The mapping data are a refinement of earlier findings, wherein radioactive in situ hybridization was used and the assignments included both the short and long arms. Results of the present study clearly exclude the short arm as the location for the three genes. Further, using a genomic cosmid clone, the GH gene was mapped to band 12p14. Compared to the earlier assignments, which included almost the entire short arm of the chromosome due to the use of radioactive in situ hybridization, the present FISH findings provide a band-specific localization for the gene. A modified, simpler version of the posthybridization trypsin/EDTA banding method is also presented.
应用荧光原位杂交(FISH)技术,使用基因组DNA克隆,在猪染色体上精确定位糖基磷脂酰肌醇(GPI)、结肠直肠癌(CRC)、脂蛋白酯酶(LIPE)和生长激素(GH)基因。利用基因组克隆和cDNA克隆,猪CRC基因被定位到6q12带。与CRC基因紧密连锁的GPI和LIPE基因,利用基因组λ克隆也被定位到同一带(6q12)。这些定位数据是对早期研究结果的细化,早期研究使用放射性原位杂交,定位结果包括短臂和长臂。本研究结果明确排除了短臂作为这三个基因的定位位置。此外,利用基因组黏粒克隆,GH基因被定位到12p14带。与早期定位结果相比,由于使用放射性原位杂交,早期定位几乎包括了染色体的整个短臂,而本FISH研究结果为该基因提供了带特异性定位。还介绍了一种改良的、更简单的杂交后胰蛋白酶/EDTA显带方法。