Molecular Cloning and Characterization of Stored mRNA in Cotyledons of Vigna nguiculata
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概要
- 論文の詳細を見る
From a λgt10 cDNA library constructed from the total polyadenylated RNA (poly A^+ RNA) of mature cowpea cotyledons, we selected recombinant phages that hybridized with a cDNA probe complementary to poly A^+ RNA from cotyledons collected 24 h after the onset of imbibition (cDNA-B) but that did not hybridize with a probe from cotyledons at developmental stage II (13 to 15 days after flowering; cDNA-A). cDNA inserts of two of these phages were subcloned in pUC18 plasmids and the resultant plasmids were designated pSAS5 and pSAS10, respectively. We also selected two recombinant phages that hybridized with cDNA-A but not with cDNA-B, and one of them was subcloned in pUC18 to generate pSASC1. We then characterized the pSAS10 cDNA insert as the cDNA of a putative stored mRNA of cowpea seeds and the pSASC1 insert as a reference cDNA. The RNA-blot hybridization after gel electrophoresis, with pSAS10 as probe, indicated that an essentially full-length cDNA had been cloned, and the hybrid-select translation of pSAS10 mRNA gave a 10-kDa product. pSASC1 mRNA was shown to code for a 46-kDa polypeptide, which corresponds in size to one of the major storage proteins of the cowpea. pSAS10 mRNA was detectable only in cotyledons at developmental stage III (17 to 19 days after flowering) or later, and the level of the mRNA began to decline when seeds germinated. The mRNA was also present in cowpea embryonic axes. mRNA hybridizable to pSAS10 cDNA was found in extracts of some other plants.
- 日本植物生理学会の論文
著者
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Minamikawa Takao
Departmen T Of Biology Tokyo Metropolitan University
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Ishibashi Nobuhiro
Department of Biology, Tokyo Metropolitan University
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Ishibashi Nobuhiro
Department Of Biology Tokyo Metropolitan University
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Ishibashi Nobuhiro
Departmen t of Biology, Tokyo Metropolitan University
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