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园艺学报 ›› 2006, Vol. 33 ›› Issue (5): 1075-1078.

• 研究报告 • 上一篇    下一篇

柑橘钙调蛋白cDNA的克隆及序列分析

岳海林1, 2;孟海军1;邓秀新1;彭抒昂2*   

  1. (1 华中农业大学作物遗传改良国家重点实验室, 湖北武汉430070; 2 华中农业大学园艺林学学院, 湖北武汉430070)
  • 收稿日期:2005-12-20 修回日期:2006-03-02 出版日期:2006-10-25 发布日期:2006-10-25

Cloning and Sequence Analysis of Citrus Calmodulin cDNA

Yue Hailin1, 2;Meng Haijun1;Deng Xiuxin1;Peng Shupang2*   

  1. (1National Key Laboratory of Crop Genetic Improvement, Huazhong Agriculture University, Wuhan, Hubei 430070, China;2College of Forestry and Horticulture, Huazhong Agriculture University, Wuhan, Hubei 430070, China)
  • Received:2005-12-20 Revised:2006-03-02 Online:2006-10-25 Published:2006-10-25

摘要:

以温州蜜柑(Citrus unshiu Marc. ) 幼果cDNA第1链为模板, 参照大麦钙调蛋白基因序列(Accession No. M27303) 合成5p端和3p端引物, 以PCR的方法扩增得到柑橘钙调蛋白cDNA, 将其克隆到PMD182T载体上并进行测序。序列分析表明, 柑橘钙调蛋白cDNA全长453 bp, 共编码148个氨基酸, 与大麦和大豆钙调蛋白基因的同源性均高达83%以上, 所编码氨基酸序列同源性达97%以上。以该cDNA作探针进行点杂交, 得到良好的杂交信号。

关键词: 柑橘, 钙调蛋白, cDNA克隆, 序列分析

Abstract: Using the first strand cDNA from the immature fruits of Citrus unshiu Marc. as template, we amplified and cloned the cDNA of Citrus calmodulin gene in this study. The PCR primers were designed according to the sequences of barley calmodulin gene obtained from GenBank (Accession No. M27303). The PCR product was cloned into PMD182T vector and then sequenced. Sequence analysis indicated that the cDNA of Citrus calmodulin gene contains 453 nucleotides coding for 148 amino acid residues. The nucleotide sequences and their deduced amino acid sequences of the Citrus calmodulin cDNA show an identity of above 83% and 97% , respectively, to those of barley and soybean. Southern dot blot analysis using the cloned cDNA as probe confirmed the results of sequencing analysis.

Key words: Citrus, Calmodulin, cDNA cloning, Sequence analysis