| 引用本文: | 崔 静,李太武,苏秀榕,李 晔,徐 静,周 君,应 琪,王中华,李松伟.南移养殖的刺参(Apostichopus japonicus)cDNA文库的构建及原肌球蛋白基因的研究.海洋与湖沼,2010,41(6):850-856. |
| |
|
| 摘要: |
| 以南移养殖的刺参为实验材料, 采用非均一化的 Oliga-dT 引物定向克隆技术构建了南移刺参混合组织的 cDNA 文库。对文库质量的分析结果表明, cDNA 文库的库容量为 2.2×107cfu, 重组率达95.8%, 平均插入片段长度750bp左右。 挑取cDNA克隆进行5’端测序, 成功测序185个反应, 分析得到 136 个单基因簇(Unigene), 其中包括 40 个重叠群(Contigs)、96 个单拷贝 EST (Singletons)。结果表明, 克隆南移养殖的刺参原肌球蛋白(tropomyosin, Trp) cDNA 全长序列为 1112bp, ORF 编码284 个氨基酸, 其预测蛋白的分子量为 33.27kDa, 等电点为 4.56。该氨基酸序列与紫石海胆、南极大磷虾、锯缘青蟹、文昌鱼、河蟹、家蚕同源性分别为 62%、51%、46%、49%、47%、46%。
|
| 关键词: 刺参, cDNA 文库, 基因克隆, 原肌球蛋白基因 |
| DOI：10.11693/hyhz201006008008 |
| 分类号: |
| 基金项目:国家农业科技成果转化资金, 2007GB2C220359 号; 浙江省重大科技专项(优先主题)重大农业项目, 2008C02009-Ⅱ-2 号; 宁波市农业科技成果转化资金, 2007C30001 号 |
附件 |
|
| CONSTRUCTION OF cDNA LIBRARY WITH APOSTICHOPUS JAPONICUS CULTURED IN THE SOUTH AND SEQUENCE ANALYSIS OF ITS TROPOMYOSIN GENE |
|
CUI Jing1, LI Tai-Wu2, SU Xiu-Rong1, LI Ye1, XU Jing3, ZHOU Jun1, YING Qi1, WANG Zhong-Hua1, LI Song-Wei1
|
|
1.Faculty of Life Science and Biotechnology, Ningbo University;2.Ningbo City College of Vocational Technology;3.Wenzhou Vocational & Technology College
|
| Abstract: |
| Based on the construction technology of non-normalized cDNA library, the cDNA libraries of Apostichopus japonicus cultured in the south was constructed. This library reached 2.2×107 in capacity; the percentage of recombination was 95.8%. PCR results showed that the average size of inserts was larger than 750bp. A total of 185 successful sequencing reactions yielded. In addition, 136 non-repetitive sequences were formed including 40 contigs and 96 singletons from these ESTs after initial assembly. The tropomyosin gene of A. japonicus, which was 1112bp long, its ORF encoding 284 amino acids with a predicted molecular weight of 33.27kDa and a theoretical isoelectric point of 4.56. The amino acid sequence was compared with Strongylocentrotus purpuratus, Euphausia superba, Portunus sanguinolentus, Branchiostoma belcheri, Eriocheir sinensis, and Bombyx mori; and high similarities of 62%—46% were revealed.
|
| Key words: Apostichopus japonicus, cDNA library, Gene cloning, Tropomyosin gene |
