Hall L, Davies M S, Craig R K
Nucleic Acids Res. 1981 Jan 10;9(1):65-84. doi: 10.1093/nar/9.1.65.
We describe the cloning of double-stranded cDNA synthesized from lactating human mammary gland total poly(A)-containing RNA, into the EcoRI site of the plasmid pAT153. Nine recombinants were shown to contain alpha-lactalbumin cDNA sequences as determined by positive hybridisation translation of complementary RNA. Restriction enzyme maps were determined for six of these. Alignment of the restriction map with the known amino acid sequence of human alpha-lactalbumin provided evidence that two plasmids, designated phO-53 and phB-35, contained the complete coding sequence of the primary translation product (pre-alpha-lactalbumin). Hybridisation studies using purified human, monkey and guinea-pig alpha-lactalbumin cDNA demonstrated that greater nucleotide sequence divergence has occurred within the rodents than the primates, and that rodent alpha-lactalbumin mRNAs retain regions of homology with primate alpha-lactalbumin mRNAs.
我们描述了从哺乳期人乳腺含总多聚腺苷酸（poly(A)）的RNA合成的双链cDNA克隆到质粒pAT153的EcoRI位点的过程。通过互补RNA的阳性杂交翻译确定，九个重组体含有α-乳白蛋白cDNA序列。对其中六个进行了限制性酶切图谱分析。将限制性图谱与已知的人α-乳白蛋白氨基酸序列进行比对，结果表明，两个命名为phO-53和phB-35的质粒包含初级翻译产物（前α-乳白蛋白）的完整编码序列。使用纯化的人、猴和豚鼠α-乳白蛋白cDNA进行的杂交研究表明，啮齿动物内部发生的核苷酸序列差异比灵长类动物更大，并且啮齿动物α-乳白蛋白mRNA保留了与灵长类动物α-乳白蛋白mRNA的同源区域。
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