| 引用本文: | 应成琦,蔡春尔,尹顺吉,徐 韧,LIN Sen-Jie,周志刚,马家海,何培民.长石莼(缘管浒苔)(Ulva linza) rbcL全长基因的克隆与序列分析.海洋与湖沼,2010,41(4):555-562. |
| |
|
| |
|
|
| 本文已被：浏览 3030次 下载 3041次 |
码上扫一扫！ |
|
|
| 长石莼(缘管浒苔)(Ulva linza) rbcL全长基因的克隆与序列分析 |
|
应成琦1, 蔡春尔1,2, 尹顺吉1, 徐 韧3, LIN Sen-Jie4, 周志刚1, 马家海1, 何培民1
|
|
1.上海海洋大学 农业部水产种质资源与养殖生态重点开放实验室;2.中国水产科学研究院东海水产研究所 中国水产科学研究院盐碱地渔业工程技术中心;3.国家海洋局东海环境监测中心;4.Department of Marine Sciences, University of Connecticut, Groton
|
|
| 摘要: |
| 克隆分离了长石莼(缘管浒苔)光合作用第一关键酶 Rubisco 大亚基全长基因(rbcL)。首先应用 PCR 和 RT-PCR 方法扩增 rbcL 大片段 DNA 序列和大片段 cDNA 序列, 结果表明, 获得的 2 个克隆序列完全相同, 该 rbcL 大片段基因不存在内含子。其次应用基因步移方法分别对 rbcL 基因的 5′上游未知序列和 3′下游未知序列进行扩增, 在此基础上, 将 3 个片段序列进行拼接, 获得了rbcL 全长基因序列(NCBI 登陆号: DQ813497)。序列全长 2124bp, 其中编码区序列长 1425bp, 为单外显子基因, 编码 474 个氨基酸; 5′非翻译区序列长 225bp, 转录起始位点为位于翻译起始密码子 ATG 上游第47 个核苷酸 G, 在距离转录起始位点?9bp—?48bp 的范围内有推测的类似原核生物的启动子序列(?10 区: TAAAAT、?35 区: TTGAAA); 3′非翻译区序列长 474bp, 在距离转译终止密码子 TAA 下游54—98bp 处有一段较长的回文序列, 可形成一个 22bp 的茎结构。密码子偏好性分析结果表明, 长石莼(缘管浒苔)rbcL 基因偏向使用第三位核苷酸碱基为 A 和 T 的密码子。
|
| 关键词: rbcL, 长石莼(缘管浒苔), 全长序列, 基因步移 |
| DOI：10.11693/hyhz201004014014 |
| 分类号: |
| 基金项目:国家自然科学基金项目, 30371101 号; 教育部博士点基金资助项目, 2006—2007; 上海市优秀学科带头人计划项目, 08XD14037 号; 上海市科委 “长三角联合攻关” 项目, 062358101 号; 上海市教委优势(重点)学科资助项目, S30701 号 |
附件 |
|
| CLONING AND SEQUENCE ANALYSIS OF THE FULL-LENGTH rbcL GENE FROM ULVA LINZA |
|
YING Cheng-Qi1,2, CAI Chun-Er1, YIN Shun-Ji1, XU Ren3, LIN Sen-Jie4, ZHOU Zhi-Gang1, MA Jia-Hai1, HE Pei-Min1
|
|
1.Aquatic Genetic Resources & Aquacultural Ecology Key Lab of the Ministry of Agriculture, Shanghai Ocean University;2.Research Center for Saline Fisheries Technology, East China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences;3.East Sea Department of State Oceanic Administration;4.Department of Marine Sciences, University of Connecticut, Groton
|
| Abstract: |
| The full-length DNA sequence of Rubisco, the first key enzyme of photosynthesis in Ulva linza was cloned. Firstly, partial rbcL DNA and cDNA sequence both with 1101bp in size were obtained using PCR and RT-PCR technology, respectively, and both of their PCR products were sequenced and analyzed. The partial rbcL DNA sequence was the same as the rbcL cDNA sequence, indicating there is no intron in the partial rbcL DNA sequence. In addition, gene walking technology was used to clone the 5′ upstream sequence and the 3′ downstream sequence. The sizes of the cloned sequences at 5′ upstream and 3′ downstream were 474bp and 1053bp, respectively. Based on these three cloned sequences, a full-length DNA sequence of 2124bp was acquired (NCBI accession number: DQ813497), which includes a coding sequence of 1425bp, encoding 474 amino acids. The 5′ upstream sequence is 225bp and includes a putative promoter between ?56bp—?95bp upstream the ATG codon. The 474bp long downstream sequence contains a long palindrome sequence locating between 54—98bp downstream from the termination codon and being capable of forming a 22bp stem. Codon preference analysis performed with the SMS software indicates that codons with the third base being A or T are in preference in the rbcL gene in Ulva linza.
|
| Key words: rbcL, Ulva linza, Full-length sequence, Gene walking |
|
|
|
|
|