| 摘要: |
| 采用RT-PCR 的方法从酵母中成功地得到了磷酸甘油酸变位酶的cDNA 基因, 分别用32P 和地高辛-ddUTP 标记以用作探针。以32P 标记的探针筛选三角褐指藻基因组文库, 获得了4kb 的阳性DNA 片段; 进一步分析发现, 该4kb 片段的真正阳性区域是位于片段端部的306bp 的序列, 因此认为该序列为三角褐指藻磷酸甘油酸变位酶基因的侧翼部分。克隆该306bp 的DNA 片段, 并且测定其序列。结果表明, 该306bp 的DNA 片段包含两个同向重复序列, 每个重复序列的大小为116bp, 在每个重复序列中均含有GGTTCAATGT 区域, 这与一般常见的真核基因5ˊ端的CAAT box 有相似之处。 |
| 关键词: 三角褐指藻, 基因组文库, 磷酸甘油酸变位酶基因, 侧翼序列 |
| DOI： |
| 分类号: |
| 基金项目:国家自然科学基金资助项目, 30170499 号; 中国科学院创新课题资助, L008 号 |
|
| SCREENING, CLONING AND SEQUENCING FOR THE PUTATIVE FLANKING REGION OF GENE ENCODING FOR PHOSPHOGLYCERATE MUTASE OF PHAEODACTYLUM TRICORNUTUM |
|
WANG Guang-Ce, SUN Hai-Bao, ZENG Cheng-Kui( C.K. Tseng)
|
|
Institute of Oceanology, The Chinese Academy of Sciences
|
| Abstract: |
| The phosphoglyceralde mutase cDNA gene, which is involved in glycolysis and catalyzes the inter-conversion of 3-phosphoglycerate and 2-phosphoglycerate, was obtained with RT-PCR from Saccharomyces cerevisiae. The gene was labeled with both radio 32P and DIG-ddUTP as probe to screen the genomic library of Phaeodactylum tricornutum. a 4kb positive DNA fragment was discovered from positive clone screened. Further analysis elucidated that the real positive region in the 4kb DNA fragment was 306bp located on the end. Therefore, the 306bp is considered to be flanking region of phosphoglycerade gene from Phaeodactylum tricornutum. The 306bp of DNA fragment was cloned into pBluescript SK+ plsmid and sequenced. The sequence result shows that the 306bp include two repeat regions with same direction whose size is 116bp, and GGTTCAATGT region which is similar to CAAT BOX in 5ˊ end of eukaryotic gene. |
| Key words: Phaeodactylum tricornutum, Genomic library, Phosphoglyceralde mutase gene, Flanking region |
