Mattei M G, Weil D, Pribula-Conway D, Bernard M P, Passage E, Van Cong N, Timpl R, Chu M L
Centre de Génétique Médicale, INSERM U242, Marseille, France.
Hum Genet. 1988 Jul;79(3):235-41. doi: 10.1007/BF00366243.
A laminin B2 chain cDNA clone was isolated from a human lung cDNA library by screening with antibody against mouse laminin. The authenticity of the human cDNA clone was established by comparison of the nucleotide and deduced amino acid sequences of the cDNA insert with those of the previously reported mouse laminin cDNA clones. The human clone (LC7) contained an insert of 0.75 kb (kilobase pair) that corresponded to the last 232 amino acid residues in the carboxyl terminus of the B2 chain. Northern blot analyses with the LC7 probe detected two mRNA transcripts of 8.2 and 5.6 kb in both normal human skin fibroblasts and three human tumor cell lines. The cDNA probe was also used in Southern blot analysis of DNA from human rodent somatic cell hybrids to localize the gene to human chromosome 1. In situ hybridization of the cDNA with metaphase chromosome spreads confirmed the assignment and further mapped the human laminin B2 chain gene to the long arm of chromosome 1 in the band q31.
通过用抗小鼠层粘连蛋白抗体筛选,从人肺cDNA文库中分离出一个层粘连蛋白B2链cDNA克隆。通过比较该cDNA插入片段的核苷酸和推导的氨基酸序列与先前报道的小鼠层粘连蛋白cDNA克隆的序列,确定了该人cDNA克隆的真实性。人克隆(LC7)包含一个0.75 kb(千碱基对)的插入片段,对应于B2链羧基末端的最后232个氨基酸残基。用LC7探针进行的Northern印迹分析在正常人皮肤成纤维细胞和三种人肿瘤细胞系中检测到8.2和5.6 kb的两种mRNA转录本。该cDNA探针还用于对人-啮齿动物体细胞杂种的DNA进行Southern印迹分析,以将该基因定位到人类1号染色体上。该cDNA与中期染色体铺片的原位杂交证实了该定位,并进一步将人层粘连蛋白B2链基因定位到1号染色体长臂的q31带。