CLONING AND SEQUENCE-ANALYSIS OF A CDNA-ENCODING RAT PREPROCHOLECYSTOKININ
CLONING AND SEQUENCE-ANALYSIS OF A CDNA-ENCODING RAT PREPROCHOLECYSTOKININ
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DOI：
10.1073/pnas.81.3.726
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发表时间：
1984-01-01
期刊：
影响因子：
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通讯作者：
DIXON, JE
中科院分区：
文献类型：
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作者：
DESCHENES, RJ;LORENZ, LJ;DIXON, JE
从大鼠甲状腺髓样癌中提取Poly（A）RNA，该癌表现出高水平的免疫反应性胆囊收缩素（CCK）。从多聚（A）RNA合成双链c[互补]DNA，并插入pBR 322的Pst I位点。使用杂交探针d（T-C-C-A-T-C-C-A-N-C-C-C-A-T-G-T-A-G-T-C）鉴定含有CCK cDNA的细菌克隆。根据猪CCK-8的已知氨基酸序列Asp-Tyr-Met-Gly-Trp-Met-Asp-Phe-NH 2推导出该探针的序列。测定了与大鼠前胆囊收缩素原mRNA互补的cDNA的核苷酸序列。该cDNA在5“-非编码区中含有33个核苷酸，在3”-非编码区中含有199个核苷酸，并且含有345个核苷酸编码CCK的前体，其为115个氨基酸（MW，12，826）。大鼠CCK基因的检查揭示了一个建议的转录控制序列类似于TATA序列位于上游33个核苷酸从一个建议的转录起始位点。CCK-39的氨基酸序列两侧是氨基末端和羧基末端延伸。CCK mRNA的分析表明，它是apprxeq. 750个核苷酸长。大鼠脑和肠的CCK mRNA的大小似乎与癌的CCK mRNA相同。
Poly(A) RNA was isolated from a rat medullary thyroid carcinoma that exhibited high levels of immunoreactive cholecystokinin (CCK). Double-stranded c[complementary]DNA was synthesized from the poly(A) RNA and inserted into the Pst I site of pBR322. Bacterial clones containing CCK cDNA were identified using the hybridization probe d(T-C-C-A-T-C-C-A-N-C-C-C-A-T-G-T-A-G-T-C). The sequence of the probe was deduced from the known amino acid sequence of porcine CCK-8, Asp-Tyr-Met-Gly-Trp-Met-Asp-Phe-NH2. The nucleotide sequence of the cDNA complementary to the mRNA of rat preprocholecystokinin was determined. The cDNA contains 33 nucleotides in the 5''-noncoding region, 199 nucleotides in the 3''-noncoding region, and 345 nucleotides coding for a precursor to CCK, which is 115 amino acids (MW, 12,826). Examination of the rat CCK gene revealed a suggested transcriptional control sequence analogous to the TATA sequence located 33 nucleotides upstream from a proposed transcriptional start site. The amino acid sequence of CCK-39 is flanked by both amino-terminal and carboxyl-terminal extensions. Analysis of CCK mRNA showed that it is .apprxeq. 750 nucleotides long. CCK mRNA of the rat brain and intestine appeared to be identical in size to the CCK mRNA of the carcinoma.