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野生大豆Δ′-吡咯琳-5-羧酸合成酶(P5CS)基因的克隆与序列分析 被引量:10

Molecular Cloning and Sequence Analysis of Δ′-pyrroline-5-carboxylate Synthetase(P5CS) Gene in Wild Soybean
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摘要 根据栽培大豆(Glycine max)Δ′-吡咯琳-5-羧酸合成酶基因的全长序列设计引物,通过RT-PCR直接扩增的方法从野生大豆中克隆到了野生大豆(Glycine soja)的同源基因,命名为GsP5CS。GsP5CS最大开放阅读框为2148bp,编码含有716个氨基酸残基、分子量为77.9 kDa的蛋白,预测等电点6.21。亲疏水性分析显示,GsP5CS含有连续的亲水片断。序列分析显示,GsP5CS与栽培大豆(Glycine max)、飞蛾豆(Vigna aconitifolia)、蒺藜苜蓿(Medicagotruncatula)、紫花苜蓿(Medicago sativa)、甘蓝型油菜(Brassica napus)、拟南芥(Arabidopsis thaliana)、水稻(Oryza sati-va)、普通小麦(Triticum aestivum)等植物的Δ′-吡咯琳-5-羧酸合成酶基因高度同源,序列相似性分别为94%、87%、87%、86%;75%、74%、72%。根据野生大豆与这8种植物的P5CS基因序列相似性所建立的进化树,显示与传统的分类地位基本一致。 GsP5CS, homologous with the GmP5CS gene encoding a △′-pyrroline -5-carboxylate synthetase was cloned from wild soybean ( Glycine soja) by RT- PCR, with primers designed according to the sequence of GmP5CS. The largest open reading frame of GsPSCS gene has 2148bp in length and encoded a protein of 716 amino acid residues. The estimated molecular weight and isoelectric points of the putative protein was 77.9 kDa and 6.21 ,respectively. Hydrepathy plot analysis indicated that the GsPSCS was hydrophilic. Blast result analysis showed that GsP5CS,GmP5CS,MsP5CS,MtP5CS,AtP5CS,BnP5CS, OsP5CS, TaP5CS shared a cluster. The phylogenetic tree shows that the relationship based on the homology is consistent with the morphological and taxonomic results.
出处 《大豆科学》 CAS CSCD 北大核心 2008年第6期915-920,共6页 Soybean Science
基金 国家重点基础研究发展计划(973计划)资助项目(2007CB116205,2004CB117203-3) 国家高技术研究发展计划(863计划)资助项目(2006AA10Z120) 农业部CWRC资助项目(CWRC-20080205) 农业部野生植物保护与利用项目(2006-2008)
关键词 野生大豆 △′-吡咯琳-5-羧酸合成酶 耐盐 Wild soybean △′- pyrroline- 5- carboxylate synthetase Salt resistance
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