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Acta Agron Sin ›› 2012, Vol. 38 ›› Issue (12): 2162-2169.doi: 10.3724/SP.J.1006.2012.02162

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Molecular Cloning and Expression Analysis of CaM-Like Protein Genes (BoCML49) from Cabbage (Brassica oleracea L. var. capitata)

SONG Ming**,XU Jun-Qiang**,SUN Zi-Jian,TANG Qing-Lin,WANG Zhi-Min,WANG Xiao-Jia*   

  1. Key Laboratory of Horticulture Science for Southern Mountainous Regions, Ministry of Education / Chongqing Key Laboratory of Olericulture / College of Horticulture and Landscape Architecture, Southwest University, Chongqing 400715, China
  • Received:2012-04-18 Revised:2012-08-15 Online:2012-12-12 Published:2012-10-08
  • Contact: 王小佳, E-mail: wxj@swu.edu.cn, Tel: 023-68251093

Abstract:

Calcium sensor proteins in plants play important roles in pollen germination and pollen tube growth processes. Calmodnlin-like (CML) protein of cabbage (Brassica oleracea L. var. capitata) was identified in the process of pollen germination in cabbage line E1 by two-dimensional electrophosis of the pollen total protein. The sequence of BoCML49 fragment was 707 bp. The expression analysis by qPCR showed that the BoCML49 gene was down-regulated when pollen germinated, 550 bp and 721 bp DNA fragments were obtained by 5'-Race and 3'-Race, respectively. We obtain BoCML49 gene with a total length of 1 343 bp by splicing, its open reading frame was located in the region from 125 to 1 078 bp, and contained a 124 bp 5' untranslated region (5' UTR) and a 265 bp 3' UTR, the polyadenylation signal (AATAAA) was located at 1 222 bp. The deduced BoCML49 protein contained 317 amino acids, with a MW of 33.51 kD and pI of 6.93. The structural analysis of BoCML49 though Smart-embl showed that it contained two critical functional domain EF-hand modifs, which located at the position of 150178 and 216244 amino acid residues, at the same time the ORF might contain seven α-helixes and ten β-sheets. The phylogenetic tree indicated that the BoCML49 had close genetic relationship with AtCML49 and AtCML50. Prokaryotic expression showed that the molecular mass of BoCML49 protein was about 37.55 kD. cDNA

Key words: Cabbage, Pollen, BoCML49, Expression analysis

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