PJB-2012-230
MOLECULAR CHARACTERIZATION AND PROKARYOTIC EXPRESSION OF ORF507 STERILITY-ASSOCIATED GENE IN CHILLI PEPPER (CAPSICUM ANNUM L.) CYTOPLASMIC MALE STERILITY
MING-HUA DENG1,2†, JIN-FEN WEN3†, JIN-LONG HUO4†, HAI-SHAN ZHU2, PEI WANG4, XIONG-ZE DAI1, ZHU-QING ZHANG1, HUI ZHOU2 AND XUE-XIAO ZOU1*
Abstract
9704A is a cytoplasmic male sterility (CMS) of chilli pepper (Capsicum annuum L) that has been commercially used in chilli pepper hybrid production in China. However, little is known about the CMS-associated gene in chilli pepper 9704A.In this study, according to the sequence of relative references, a fragment of 593 bp including a 507-bp single-coding exon, Orf507 (GenBank accession No. GU357841) was cloned from leaves of chilli pepper CMS-9704A. Results revealed that the orf456-2 had one nucleotide deleted in codon 150 (nt450delC) and consists of a 507-bp single-coding exon which encodes 168 amino acid compared with the original sequence data of orf456. RT-PCR results showed that the Orf507 was transcribed in buds and leaves at all developmental stages of CMS-9704A and a hybrid F1, while no band was detected in the maintainer line. The nuclear restorer gene may restore fertility by post-transcriptional processing or translational/post-translational modification, but did not interfere with transcription of Orf507 directly. The entire coding region of the Orf507 was inserted into an expression vector pET32a(+) and transformed into Escherichia coli Rosetta. Expression of the fusion protein was successfully induced by isopropyl-β-D-thiogalactopyranoside. This study could establish a basis for understanding the molecular mechanism of chilli pepper CMS.
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