The use of a heptadeoxyribonucleotide as a specific primer for prolactin mRNA: a prediction of ambiguous RNA splicing.
The use of a heptadeoxyribonucleotide as a specific primer for prolactin mRNA: a prediction of ambiguous RNA splicing.
复制标题
使用七脱氧核糖核苷酸作为催乳素 mRNA 的特异性引物：对模糊 RNA 剪接的预测。
DOI：
10.1016/0006-291x(81)91647-8
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发表时间：
1981
影响因子：
3.1
通讯作者：
Dixon,JE
中科院分区：
文献类型：
--
作者：
Taylor,WL;Collier,KJ;Weith,HL;Dixon,JE
摘要利用大鼠垂体前叶的聚(A)+ RNA合成135个核苷酸的cDNA，利用5 '标记的寡核苷酸d （TCATCAT）与泌乳素前氨基酸序列（Met·Met）的mRNA序列互补。核苷酸序列分析表明，该cDNA与大鼠催乳素mRNA的逆转录相对应。这些结果表明，短如七聚体的寡核苷酸在其5 '端有单核苷酸错配，可以用来获得可直接测序的特定cDNA。寡核苷酸诱导的cDNA序列和克隆的大鼠催乳素mRNA序列[Cooke， N., et al.， J. Biol.]。Chem. 255, 6502(1980)]的不同之处在于，前者序列在大鼠泌乳素前信号序列的第10个氨基酸对应的位置上有一个额外的丙氨酸密码子（GCA）。这一结果可以解释为在前体RNA的外显子-内含子连接处产生的两个大鼠催乳素mRNA群体。
Abstract A 5′-labelled oligonucleotide, d (TCATCAT), complementary to the mRNA sequence coding for the amino acid sequence (Met· Met) present in preprolactin was used to initiate the synthesis of a 135 nucleotide cDNA from poly (A)+ RNA isolated from the rat anterior pituitary. Nucleotide sequence analysis of the cDNA revealed that it corresponded to a reverse transcript of rat prolactin mRNA. These results established that an oligonucleotide as short as a heptamer with a single nucleotide mismatch at its 5′ end can be used to obtain a specific cDNA which can be sequenced directly. The oligonucleotide initiated cDNA sequence and the sequence of a cloned cDNA to rat prolactin mRNA [Cooke, N., et al., J. Biol. Chem. 255, 6502 (1980)] differ in that the former sequence has an additional codon (GCA) for alanine at a position corresponding to the tenth amino acid of the signal sequence of rat preprolactin. This result could be explained by two populations of rat prolactin mRNA arising from ambiguous splicing at an exon-intron junction in the precursor RNA.