《植物生理学报》 2010, 46(1): 37-41

银杏叶绿体petD 基因的克隆与表达

许锋¹, 张威威¹, 唐寅¹, 王燕¹, 程水源^2,*

1 长江大学园艺园林学院, 湖北荆州434025; 2 黄冈师范学院生命科学与工程学院, 湖北黄冈438000

通信作者：程水源；E-mail: s_y_cheng@sina.com；Tel: 0713-88 335 99

摘　要：

根据黑松、云杉、菠菜与玉米叶绿体 petD 基因序列设计引物, 以银杏叶绿体基因组 DNA 为模板, PCR 扩增克隆了 银杏叶绿体 petD 基因(GenBank 登录号为 DQ923066, 命名为 GbpetD)的序列。序列分析显示, GbpetD 基因组 DNA 序列编 码区长 1 243 bp, 含 1 个内含子和 2 个外显子, 其外显子序列编码 177 个氨基酸。相似性比对显示, 该基因编码区序列与云 杉、台东苏铁、黑松、莴苣、木薯、北美落叶松的 petD 基因核苷酸同源性为 84%~99%, 氨基酸序列同源性为 85%~93%。 系统进化树分析结果表明 GbpetD 蛋白质与黑松、北美落叶松、云杉、苏铁等裸子植物的 petD 蛋白质聚类关系最近。半 定量 RT-PCR 分析表明, GbpetD 基因在银杏叶和茎中表达, 在叶中表达量最大。

关键词：银杏; GbpetD基因; 序列分析; 表达

收稿：2009-10-28   修定：2009-11-24

资助：湖北省自然科学基金(2008CDA061)、湖北省教育厅科学研究计划优秀中青年人才项目(Q20091201)和长江大学博士基金(0010 113)。

Cloning and Expression of A Chloroplast petD Gene from Ginkgo biloba L.

XU Feng¹, ZHANG Wei-Wei¹, TANG Yin¹, WANG Yan¹, CHENG Shui-Yuan^2,*

¹College of Horticulture and Garden, Yangtze University, Jingzhou, Hubei 434025, China; ²College of Life Science and Engineering, Huanggang Normal University, Huanggang, Hubei 438000, China

Corresponding author: CHENG Shui-Yuan; E-mail: s_y_cheng@sina.com; Tel: 0713-88 335 99

Abstract:

A chloroplast petD gene, named GbpetD, was cloned from Ginkgo biloba using a pair of primers, which were designed according to the homologous sequences in Pinus thunbergii, Picea abies, Spinacia oleracea, and Zea mays. The genomic DNA of GbpetD had 1 243 bp coding sequence, including one intron and two exons, which encoding a 177-amino-acid protein. The Blast results showed that the homology of the GbpetD nucleotide sequence with petD genes from P. abies, Cycas taitungensis, P. thunbergii, Lactuca sativa, Manihot esculenta, and Larix occidentalis was from 84% to 99%, and the homology of amino acid sequences was from 85% to 93%. Phylogenetic tree analysis indicated that the GbpetD protein had the closest relationship with petD proteins from gymnosperm plants. The expression analysis with semi-quantitative RT-PCR revealed that GbpetD gene expressed in leaves and young stems, and the highest level was detected in leaves.

Key words: Ginkgo biloba; GbpetD; sequence analysis; expression