Breslow J L, Ross D, McPherson J, Williams H, Kurnit D, Nussbaum A L, Karathanasis S K, Zannis V I
Proc Natl Acad Sci U S A. 1982 Nov;79(22):6861-5. doi: 10.1073/pnas.79.22.6861.
We have isolated cDNA clones encoding human apolipoprotein (apo) A-I. Twenty putative apo A-I cDNA clones were selected by screening 10,000 clones of an adult human liver cDNA library with an oligonucleotide probe. The probe was a mixture of synthetic 14-base-long DNA oligomers constructed to correspond to the codons for apo A-I amino acids 105-109. Four of these clones were examined further and showed 600- to 800-base-pair (bp) inserts. Preliminary restriction mapping and partial DNA sequence analysis indicated that the shorter inserts were a subset of the longer DNA inserts. DNA sequence analysis of the clone with an insert of approximately equal to 600 bp, designated pAI-113, revealed that it contained a DNA sequence corresponding to apo A-I amino acids 94-243. The DNA base sequence of this clone also contained a standard termination codon, polyadenylylation signal, and poly(A) tail. Partial DNA sequence of a second clone that contained an 800-bp insert, designated pAI-107, showed that it corresponded to apo A-I amino acids 18-243 and also included the 3' untranslated region. Isolation of these cDNA clones will facilitate molecular analyses of apolipoproteins in normal and disease states.
我们已经分离出编码人载脂蛋白(apo)A-I的cDNA克隆。通过用寡核苷酸探针筛选10000个成人肝脏cDNA文库克隆,挑选出20个推定的apo A-I cDNA克隆。该探针是由合成的14个碱基长的DNA寡聚物混合而成,其构建对应于apo A-I氨基酸105 - 109的密码子。对其中4个克隆进行了进一步检测,发现插入片段为600至800碱基对(bp)。初步的限制性图谱分析和部分DNA序列分析表明,较短的插入片段是较长DNA插入片段的一个子集。对插入片段约600 bp的克隆(命名为pAI - 113)进行DNA序列分析,结果显示它包含对应于apo A-I氨基酸94 - 243的DNA序列。该克隆的DNA碱基序列还包含一个标准终止密码子、聚腺苷酸化信号和聚(A)尾。对第二个包含800 bp插入片段的克隆(命名为pAI - 107)进行的部分DNA序列分析表明,它对应于apo A-I氨基酸18 - 243,并且还包括3'非翻译区。这些cDNA克隆的分离将有助于对正常和疾病状态下的载脂蛋白进行分子分析。