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ACTA HORTICULTURAE SINICA ›› 2012, Vol. 39 ›› Issue (4): 677-686.

• Vegetables • Previous Articles     Next Articles

Cloning and Expression Analysis of Calmodulin Gene in Brassica oleracea L. var. capitata L.

SUN  Zi-Jian, WEI  Jing-Yi, WANG  Xiao-Jia, SONG  Ming, TANG  Qing-Lin, WANG  Zhi-Min, REN  Xue-Song   

  1. (College of Horticulture and Landscape Architecture,Southwest University;Chongqing Key Laboratory of Olericulture;Key Laboratory of Horticulture Science for Southern Mountainous Regions,Ministry of Education,Chongqing 400715,China)
  • Online:2012-04-25 Published:2012-04-25

Abstract: Calmodulin(CaM)mediates lots of cellular processes in eukaryotes,it is a pivotal transducer of calcium signal. A Brassica oleracea CaM gene was isolated using homology based on the cloning method. The ORF(open reading frame)of this gene was 450 bp encoding 149 amino acids with  four conserved EF-hand domains. There is no transmembrane domain or signal peptide sequence in tentative protein sequence. The recombinant CaM proteins(three mutants included)were expressed in E. coli,and subsequently separated on SDS-PAGE in the presence of EGTA,their electrophoresis motilities were related with that of mutant EF-Hand motifs. The activity assay showed that the CaM activity relied on the presence of Ca2+. Gene expression in the pollen germination was investigated using qPCR,the results indicated that the gene expression increased first and then descended in pollen of self-incompatible line,but decreased continuously in self-compatible line,both of which were down-regulated significantlywhen TFP was added in pollens;There was no significant difference in the beginning or at the end of pollen germination when W-7 sepharose was added.

Key words: cabbage, pollen, calmodulin, cloning, expression analysis

CLC Number: