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Cloning and expression of PFP #alpha# subunit cDNA in rice

Wang Zhaoming, Wang Yuehao, Gao Min, Shi Jiaonai

Shanghai Institute of Plant Physiology,The Chinese Academy of Sciences.Shanghai 200032

Abstract

Potato anti-rabbit PFP #alpha# subunit antibody was purified by affinity elution. The antigen specificity was well confirmed by Western immunoblot. 3 positive signals were obtained from 5×10~(4) pfu plaques when a rice seed cDNA library was screened with this spcific antibody. The positive clones were confirmed by further replating and rescreening. #lambda# DNA was extracted and digested with Not I-Sal I, agarose gel electrophoresis showed that the longest inserted fragment was about 2.1 kb in length. The 2.1 kb fragment has been subcloned into plasmid PSK. Southern blot analysis reveals the homogeneity between the cloned cDNA and potato PFP #alpha# subunit cDNA probe. We got a peptide product of 66 kD by in vitro transcription and translation of the cloned gene.

Key words: PFP (pyrophosphate: D-fructose-6-phosphate 1-phosphotransferase);#alpha# subunit;cDNAclone;Immunological screen

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Citing This Article：

Wang Zhaoming, Wang Yuehao, Gao Min, Shi Jiaonai. Cloning and expression of PFP #alpha# subunit cDNA in rice. Acta Physiol Sin 1997; 49 (3): (in Chinese with English abstract).