Purification of the mRNA for chicken very low density lipoproteinII and molecular cloning of its full-length double-stranded cDNA.
Purification of the mRNA for chicken very low density lipoproteinII and molecular cloning of its full-length double-stranded cDNA.
复制标题
鸡极低密度脂蛋白II mRNA的纯化及其全长双链cDNA的分子克隆。
DOI：
10.1093/nar/7.8.2147
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发表时间：
1979
影响因子：
14.9
通讯作者：
M. Gruber
中科院分区：
文献类型：
--
作者：
B. Wieringa;W. Roskam;A. Arnberg;J. van der Zwaag;G. Ab;M. Gruber
The mRNA coding for the small apo-Very Low Density Lipoprotein (apo-VLDLII) from chicken serum was highly enriched by oligo(dT) chromatography and preparative gel electrophoresis of estrogenised liver RNA. Double-stranded cDNA was synthesised by the subsequent actions of reverse transcriptase and DNA polymerase, and used for a preliminary characterisation of the structural gene. Molecular cloning of dC-tailed ds-cDNA into the Pst I site of plasmid pBR 322 yielded several recombinant clones. Five chimeric DNAs were selected and characterised by restriction enzyme mapping and electron microscopy of R-loops. At least two of them (pVLDLII 3.33 and pVLDLII 4.82) contain an almost full-length ds-transcript of VLDLII mRNA in which no more than 10-20 bases at the 5'- end are missing.
