| 桃叶片β- 半乳糖苷酶基因全长cDNA克隆及原核表达 |
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| 引用本文： | 卞伟华,江昌俊,王朝霞. 桃叶片β- 半乳糖苷酶基因全长cDNA克隆及原核表达[J]. 园艺学报, 2006, 33(4): 721-724 |
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| 作者姓名： | 卞伟华 江昌俊 王朝霞 |
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| 作者单位： | (安徽农业大学茶叶生物化学与生物技术重点实验室, 安徽合肥230036; 安徽农业大学生物技术中心, 安徽合肥230036; 安徽教育学院生物系, 安徽合肥230061; 山东滨州医学院生化教研室, 山东滨州256603) |
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| 基金项目： | 安徽省教育厅自然科学基金 |
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| 摘 要： | 利用RT-PCR和RACE技术, 从蟠桃(Amygdalus persica var. compressa Bean) 中克隆出3 285 bp的β - 半乳糖苷酶基因cDNA全长, GenBank登录号为AY874412。该cDNA 2 559 bp, 编码853个氨基酸。将其插入到大肠杆菌表达载体pET-32a ( + ) 中, 转化BL21 trxB (DE3) , 筛选重组菌株。经IPTG诱导表达后, SDS-PAGE呈现约115 kDa特异表达条带。并通过体外酶促反应分析表达的融合蛋白的生物活性。
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| 关 键 词： | 桃 β - 半乳糖苷酶 cDNA末端快速扩增 cDNA克隆 原核表达 |
| 文章编号： | 0513-353X（2006）04-0721-04 |
| 收稿时间： | 2005-06-18 |
| 修稿时间： | 2005-06-182005-10-11 |
Cloning and Expression of β-galactosidase Gene cDNA in Peach |
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| Bian Weihua,Jiang Changjun,Wang Zhaoxia. Cloning and Expression of β-galactosidase Gene cDNA in Peach[J]. Acta Horticulturae Sinica, 2006, 33(4): 721-724 |
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| Authors: | Bian Weihua Jiang Changjun Wang Zhaoxia |
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| Affiliation: | ( Key Laboratory of Tea Biochemistry & Biotechnology, Anhui Agricultural University, Hefei, Anhui 230036, China;Biotechnology Center, Anhui Agricultural University, Hefei, Anhui 230036, China; Department of Biology, Anhui Institute of education, Hefei, Anhui 230061, China; Department of Biochemistry, Binzhou Medical College, Binzhou, Shandong 256603, China) |
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| Abstract: | A 3 285 bp cDNA encoding β-galactosidase was cloned from compress peach by the methods of RT-PCR and rapid amplification of cDNA ends (RACE). The accession number is AY874412, which contains 2 559 bp, encoding a 853 residue polypeptide. The cDNA was constructed into the vector pET-32a( + ) designed for recombinant strain. After IPTG induction, it was observed there was a special band about 115 kDa on SDS-PAGE. The activity of the fusion protein was analyzed by in vitro enzymatic reaction. |
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| Keywords: | Peach β-galactosidase Rapid amplification of cDNA ends cDNA cloning Prokaryotic expression |
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