Identification of cDNA clones encoding vasolin-containing protein and other plant plasma membrane-associated proteins by a general immuno-screening strategy.

An approach was developed for the isolation and characterization of soybean plasma membrane-associated proteins by immunoscreening of a cDNA expression library. An antiserum was raised against purified plasma membrane vesicles. In a differential screening of approximately 500,000 plaque-forming units with the anti-(plasma membrane) serum and DNA probes derived from highly abundant clones isolated in a preliminary screening, 261 clones were selected from approximately 1,200 antiserum-positive plaques. These clones were classified into 40 groups by hybridization analysis and 5'- and 3'-terminal sequencing. By searching nucleic acid and protein sequence data bases, 11 groups of cDNAs were identified, among which valosin-containing protein (VCP), clathrin heavy chain, phospholipase C, and S-adenosylmethionine:delta 24-sterol-C-methyltransferase have not to date been cloned from plants. The remaining 29 groups did not match any current data base entries and may, therefore, represent additional or yet uncharacterized genes. A full-length cDNA encoding the soybean VCP was sequenced. The high level of amino acid identity with vertebrate VCP and yeast CDC48 protein indicates that the soybean protein is a plant homolog of vertebrate VCP and yeast CDC48 protein.

Dixon, Richard A.Samuel Roberts Noble Foundation (författare)
Gonzales, Robert A.Samuel Roberts Noble Foundation (författare)
Kjellbom, PerLund University,Lunds universitet,Biokemi och Strukturbiologi,Centrum för Molekylär Proteinvetenskap,Kemiska institutionen,Institutioner vid LTH,Lunds Tekniska Högskola,Biochemistry and Structural Biology,Center for Molecular Protein Science,Department of Chemistry,Departments at LTH,Faculty of Engineering, LTH(Swepub:lu)pbio-pkj (författare)
Bhattacharyya, Madan K.Samuel Roberts Noble Foundation (författare)
Samuel Roberts Noble FoundationBiokemi och Strukturbiologi (creator_code:org_t)

Ingår i:Proceedings of the National Academy of Sciences: Proceedings of the National Academy of Sciences92, s. 4457-44611091-64900027-8424

Av författaren/redakt...: Shi, Jinrui; Dixon, Richard A ...; Gonzales, Robert ...; Kjellbom, Per; Bhattacharyya, M ...

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