| XUAN Jinsong,FENG Yanbin,WENG Manli,ZHAO Ge,SHI Jinfeng,YAO Jianting,WANG Xiuliang,GUO Baotai,QIAO Lixian,DUAN Delin,WANG Bin. 2012. Expressed sequence tag analysis and cloning of trehalose-6-phosphate synthase gene from marine alga Laminaria japonica (Phaeophyta). Acta Oceanologica Sinica, (6):139－148 |
| Expressed sequence tag analysis and cloning of trehalose-6-phosphate synthase gene from marine alga Laminaria japonica (Phaeophyta) |
| Expressed sequence tag analysis and cloning of trehalose-6-phosphate synthase gene from marine alga Laminaria japonica (Phaeophyta) |
| Received:May 05, 2011 Revised:November 14, 2011 |
| DOI：10.1007/s13131-012-0260-6 |
| Key words:cDNA library comparative study expressed sequence tag (EST) analysis Laminaria japonica trehalose-6-phosphate synthase (TPS) TPS gene |
| 中文关键词: cDNA library comparative study expressed sequence tag (EST) analysis Laminaria japonica trehalose-6-phosphate synthase (TPS) TPS gene |
| 基金项目:The National Natural Science Foundation of China under contract No. 043113159; the State Key Project in Development of New Plant Varieties by Gene Transformation under contract No. 2009ZX08009-100B; Shandong Agricultural Seedstock Breeding Project. |
| Author Name | Affiliation | E-mail | | XUAN Jinsong | The State Key Laboratory of Plant Genomics, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China
Department of Biological Science and Engineering, School of Chemical and Biological Engineering, University of Science and Technology Beijing, Beijing 100083, China | | | FENG Yanbin | The State Key Laboratory of Plant Genomics, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China | | | WENG Manli | The State Key Laboratory of Plant Genomics, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China | | | ZHAO Ge | The State Key Laboratory of Plant Genomics, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China | | | SHI Jinfeng | The State Key Laboratory of Plant Genomics, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China | | | YAO Jianting | The State Key Laboratory of Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China | | | WANG Xiuliang | The State Key Laboratory of Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China | | | GUO Baotai | College of Life Sciences, Qingdao Agricultural University, Qingdao 266109, China | | | QIAO Lixian | College of Life Sciences, Qingdao Agricultural University, Qingdao 266109, China | | | DUAN Delin | The State Key Laboratory of Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China | | | WANG Bin | The State Key Laboratory of Plant Genomics, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China | bwang@genetics.ac.cn |
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| Abstract: |
| A high quality cDNA library was constructed from the brown alga Laminaria japonica, with the titer of 1.2×105 pfu/ml. The average insert size of the cDNA library is about 1.6 kb. From the cDNA library, 591 cDNA clones were randomly selected and sequenced. As a result, 574 EST (expressed sequence tag) sequences were generated. All of the 574 ESTs were submitted to the dbEST database section of GenBank with the accession numbers from CX942625 to CX943198. The cDNA library was screened with a α-32p labeled 453 bp TPS gene probe, which is a partial sequence yielded from Porphyra yezoensis. Four positive cDNA clones were screened and the sequencing data showed that these four cDNA clones covered majority of L. japonica TPS cDNA sequence. After PCR amplification, sequencing and assembling, the entire ORF (open reading frame) sequence of the TPS gene was obtained, which was named LjTPS. LjTPS encodes a protein containing 908 amino acids with a calculated molecular mass of 101 674 Daltons. The LjTPS gene was successfully expressed in E. coli and rice. The LjTPS gene has potential application both in plant breeding to stress tolerance and in deciphering the TPS gene function and mechanism to stress tolerance. |
| 中文摘要: |
| A high quality cDNA library was constructed from the brown alga Laminaria japonica, with the titer of 1.2×105 pfu/ml. The average insert size of the cDNA library is about 1.6 kb. From the cDNA library, 591 cDNA clones were randomly selected and sequenced. As a result, 574 EST (expressed sequence tag) sequences were generated. All of the 574 ESTs were submitted to the dbEST database section of GenBank with the accession numbers from CX942625 to CX943198. The cDNA library was screened with a α-32p labeled 453 bp TPS gene probe, which is a partial sequence yielded from Porphyra yezoensis. Four positive cDNA clones were screened and the sequencing data showed that these four cDNA clones covered majority of L. japonica TPS cDNA sequence. After PCR amplification, sequencing and assembling, the entire ORF (open reading frame) sequence of the TPS gene was obtained, which was named LjTPS. LjTPS encodes a protein containing 908 amino acids with a calculated molecular mass of 101 674 Daltons. The LjTPS gene was successfully expressed in E. coli and rice. The LjTPS gene has potential application both in plant breeding to stress tolerance and in deciphering the TPS gene function and mechanism to stress tolerance. |
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