ISOLATION AND CHARACTERIZATION OF CDNA CLONES FOR HUMAN APOLIPOPROTEIN A-I
ISOLATION AND CHARACTERIZATION OF CDNA CLONES FOR HUMAN APOLIPOPROTEIN A-I
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DOI：
10.1073/pnas.79.22.6861
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发表时间：
1982-01-01
期刊：
影响因子：
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通讯作者：
ZANNIS, VI
中科院分区：
文献类型：
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作者：
BRESLOW, JL;ROSS, D;ZANNIS, VI
分离编码人载脂蛋白（apo）A-I的cDNA [互补DNA]克隆。通过用寡核苷酸探针筛选成人肝cDNA文库的10，000个克隆，选择20个推定的apo A-I cDNA克隆。探针是合成的14个碱基长的DNA寡聚体的混合物，构建为对应于apo A-I氨基酸105-109的密码子。这些克隆中的四个进一步检查，并显示600至800个碱基对（bp）的插入。初步的酶切图谱和部分DNA序列分析表明，较短的插入片段是较长的DNA插入片段的一个子集。具有插入片段的克隆的DNA序列分析命名为派-113的600 bp的DNA序列表明，它含有对应于apo A-I氨基酸92-243的DNA序列。该克隆的DNA碱基序列还含有标准终止密码子、多聚腺苷酸化信号和poly（A）尾。含有800-bp插入片段的第二个克隆（命名为派-107）的部分DNA序列表明，它对应于apo A-I氨基酸18-243，还包括3“非翻译区。这些cDNA克隆的分离将促进正常和疾病中载脂蛋白的分子分析[例如，丹吉尔病]国家。
cDNA [complementary DNA] clones were isolated 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 .apprxeq. 600 bp, designated pAI-113, revealed that it contained a DNA sequence corresponding to apo A-I amino acids 92-243. 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 [e.g., Tangier disease] states.