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克隆和表达分析 DNA 序列的主要乳蛋白的罂粟。

Cloning and expression analysis of DNA sequences for the major latex protein of opium poppy.

机构信息

Department of Biology, Texas A & M University, 77843-3258, College Station, TX, USA.

出版信息

Planta. 1990 Mar;180(4):487-91. doi: 10.1007/BF02411445.

Abstract

Opium-poppy (Papaver somniferum L.) latex contains a group of very abundant, laticifer-specific peptides called the major latex proteins (MLPs). We determined a partial amino-acid sequence of an MLP cyanogen bromide peptide fragment that was used to design an MLP oligonucleotide primer. An MLP-specific DNA probe was then generated by polymerase chain reaction (PCR) of first-strand complementary DNA (cDNA) templates primed with the MLP oligonucleotide and oligodeoxythymidylic acid. This DNA fragment, called MLP-PCR, and two partial MLP cDNAs isolated with it, all contain open reading frames matching the known MLP amino-acid sequence. RNA gel blots of latex, tissue cultures, and the major organs of mature plants of opium poppy show that MLP is coded for by an 860-nucleotide mRNA and that this accumulates exclusively in laticifers.

摘要

罂粟乳胶含有一组非常丰富的乳管特异性肽,称为主要乳胶蛋白 (MLP)。我们确定了 MLP 溴化氰肽片段的部分氨基酸序列,该序列用于设计 MLP 寡核苷酸引物。然后,通过用 MLP 寡核苷酸和寡脱氧胸苷酸启动的第一链互补 DNA (cDNA) 模板的聚合酶链反应 (PCR) 生成 MLP 特异性 DNA 探针。该 DNA 片段称为 MLP-PCR,与其分离的两个部分 MLP cDNA 均包含与已知 MLP 氨基酸序列匹配的开放阅读框。乳胶、组织培养物和成熟罂粟植物主要器官的 RNA 凝胶印迹表明,MLP 由 860 个核苷酸的 mRNA 编码,并且该 mRNA 仅在乳管中积累。

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