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Cloning and sequence analysis of cDNA encoding thiamin‐binding proteins from sesame seeds
Author(s) -
Watanabe Katsumi,
Takahashi Hideyuki,
Mitsunaga Toshio
Publication year - 2001
Publication title -
physiologia plantarum
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.351
H-Index - 146
eISSN - 1399-3054
pISSN - 0031-9317
DOI - 10.1034/j.1399-3054.2001.1120412.x
Subject(s) - complementary dna , sesamum , biology , peptide sequence , microbiology and biotechnology , open reading frame , cdna library , gene , clone (java method) , cloning (programming) , sequence analysis , amino acid , genetics , nucleic acid sequence , biochemistry , computer science , horticulture , programming language
The amino acid sequences of the large polypeptides of thiamin‐binding proteins (TBPs) from sesame ( Sesamum indicum L.) seeds (STBP‐I, ‐II and ‐III) were analyzed. The large polypeptides of STBP‐I, ‐II and ‐III had the same amino acid sequences as did their small polypeptides. The peptide sequence information obtained from STBPs was used to synthesize DNA primers for amplification of the gene(s) encoding STBPs. A 200‐bp fragment was amplified from cDNA synthesized from RNA from sesame seeds 4 weeks after flowering. The 200‐bp fragment was used to clone full‐length cDNA(s) encoding STBP(s) with RACE techniques. A 644‐bp fragment was amplified, cloned and sequenced. The cDNA was a full‐length clone encoding STBP(s). It contained an open reading frame, which defined a 143‐residue polypeptide. The identified small and large polypeptide sequences of STBPs exactly matched the sequence encoded within the cDNA clone. These results indicated that the small and large polypeptides of STBPs were encoded on the mRNA as a single large proprotein precursor and that the final mature forms were generated by post‐translational processing in the same manner as the other 2S albumins of plant seeds.