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樊金萍, 柏锡, 李勇, 纪巍, 王希, 朱延明. 野生大豆SAMS同源基因的克隆及分析[J]. 植物科学学报, 2008, 26(3): 235-239.
引用本文: 樊金萍, 柏锡, 李勇, 纪巍, 王希, 朱延明. 野生大豆SAMS同源基因的克隆及分析[J]. 植物科学学报, 2008, 26(3): 235-239.
FAN Jin-Ping, BAI Xi, LI Yong, JI Wei, WANG Xi, ZHU Yan-Ming. Cloning and Analysis of Homological Genes SAMS from Glycine soja[J]. Plant Science Journal, 2008, 26(3): 235-239.
Citation: FAN Jin-Ping, BAI Xi, LI Yong, JI Wei, WANG Xi, ZHU Yan-Ming. Cloning and Analysis of Homological Genes SAMS from Glycine soja[J]. Plant Science Journal, 2008, 26(3): 235-239.

野生大豆SAMS同源基因的克隆及分析

Cloning and Analysis of Homological Genes SAMS from Glycine soja

  • 摘要: 硫腺苷甲硫氨酸作为甲基供体在转甲基反应中起到重要作用。从低温(4℃)、高盐(200 mmol/L NaCl)及干旱(30%PEG)处理的东北野生大豆幼苗叶片中提取总RNA,经反转录合成cDNA第一链,以cDNA第一链为模板,应用已知的EST序列设计一对PCR引物扩增SAM合成酶基因的全长序列,最后经BLAST比较分析,确定所获得的SAMS基因序列完整性。所得序列长1185 bp,具有完整的开放读码框,编码394个氨基酸,与棉豆和苜蓿的SAM合成酶基因的核苷酸序列的同源性分别为89%、85%。构建了以pET-32b为基础的原核表达载体并进行原核表达分析,分析SAMS基因的表达情况。为后续的抗逆性功能验证奠定基础。

     

    Abstract: S-adenosylmethionine(SAM)plays important roles in trans-methyl reactions.Under drought(30%PEG),salinity(200 mmol/L NaCl) and low temperature(4℃),total RNA was extracted from the leaf of Glycine soja and the first strand of cDNA was synthesized with reverse transcription.S-adenosylmethionine synthetase gene(SAMS gene) was amplified by PCR with the first strand cDNA as template and a pair of primers which was based on constructed ESTs sequence.Full-length SAMS gene sequence was obtained by BLAST comparison.According to the analysis,completed sequences of SAMS gene was integrality.The sequence of the SAMS gene is 1185 bp in length with an opening reading frame(ORF) encoding 394 amino acids.The cDNA sequence shows a significant homology to the SAMS genes from Phaseolus lunatus(89%),Medicago sativa(85%).There was a prokaryotic expression vectors based on pET-32b had been constructed and prokaryotic expression analyzed;To lay a strong foundation for resist adversity function analysis through situation of genic expression analysis.

     

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