Direct immunological identification of full-length cDNA clones for plant protein without gene fusion to E. coli protein.
By immunological screening of a cDNA library constructed from potato tuber poly(A)+ RNA and Escherichia coli expression vector pUC8 by the vector-primer and linker procedure of Okayama and Berg [(1982) Mol. Cell Biol. 2, 161-170], nearly full-length cDNA clones for patatin, a major protein of potato tuber, were identified. The cDNA carrying part of the 5'-noncoding region of the patatin mRNA, in addition to entire coding and 3'-noncoding regions, expressed prepatatin in E. coli cells by translational initiation inside cDNA. These results suggest that nearly full-length cDNA clones with entire coding region can be identified directly by immunological screening without gene fusion to E. coli proteins at least for some plant mRNAs.
K
K
Nakamura
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T
Hattori
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T
Asahi
D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D002265:Carboxylic Ester Hydrolases; D004247:DNA / Q000032:analysis*; D004926:Escherichia coli / Q000235:genetics; D010940:Plant Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D010957:Plasmids; D011061:Poly A / Q000378:metabolism; D014176:Protein Biosynthesis; D012313:RNA / Q000378:metabolism; D012333:RNA, Messenger
D001426:Bacterial Proteins; D010940:Plant Proteins; D012333:RNA, Messenger; C043222:patatin protein, Solanum tuberosum; D011061:Poly A; D012313:RNA; D004247:DNA; D002265:Carboxylic Ester Hydrolases