黄花苜蓿Mfβ-Hex1基因的克隆及生物信息学分析
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摘要
为了鉴定黄花苜蓿耐牧性的相关基因,采用PCR技术对黄花苜蓿Mfβ-Hex1基因进行克隆,并利用生物信息学软件分析了基因的序列信息。结果表明:黄花苜蓿β-氨基己糖苷酶(Mfβ-Hex1)基因全长786 bp,包含675 bp的开放阅读框,编码224个氨基酸,主要由谷氨酸、缬氨酸、亮氨酸等组成。与豆科植物蒺藜苜蓿和鹰嘴豆中的β-Hex1基因具有高度同源性,与蒺藜苜蓿聚为一支。Mfβ-Hex1基因属于GH20家族成员,不具有跨膜区域,其二级结构以α-螺旋为主,亚细胞定位主要存在于细胞质中,该研究为深入研究Mfβ-Hex1基因的功能奠定了基础。
In order to identify the grazing resistance related genes of Medicago falcata,the Mfβ-Hex1 gene was cloned by PCR,and the bioinformatics of the genes was analyzed by software.The results show that the β-aminohexanidase(Mfβ-Hex1) gene obtained from Medicago falcata was 786 bp,contains an open reading frame of 675 bp,and encodes 224 amino acids,mainly composed of glutamate,valine,leucine,etc.It has high homology with the Mfβ-Hex1 gene in Medicago truncatula and chickpea.Evolutionary information show that Medicago falcata is clustered with Medicago truncatula in the same group.The Mfβ-Hex1 gene belongs to the GH20 family.It doesn′ t have a transmembrane region and its secondary structure is mainly α-helix.Its subcellular localization is in the cytoplasm.This study lays a foundation for further research on the function of the Mfβ-Hex1 gene.
In order to identify the grazing resistance related genes of Medicago falcata,the Mfβ-Hex1 gene was cloned by PCR,and the bioinformatics of the genes was analyzed by software.The results show that the β-aminohexanidase(Mfβ-Hex1) gene obtained from Medicago falcata was 786 bp,contains an open reading frame of 675 bp,and encodes 224 amino acids,mainly composed of glutamate,valine,leucine,etc.It has high homology with the Mfβ-Hex1 gene in Medicago truncatula and chickpea.Evolutionary information show that Medicago falcata is clustered with Medicago truncatula in the same group.The Mfβ-Hex1 gene belongs to the GH20 family.It doesn′ t have a transmembrane region and its secondary structure is mainly α-helix.Its subcellular localization is in the cytoplasm.This study lays a foundation for further research on the function of the Mfβ-Hex1 gene.
引文
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[2]戎郁萍,韩建国,胡跃高,等.放牧苜蓿品种选育研究进展[J].中国草地学报,2004,26(1):50.
[3]赵爱奇,刘嘉荣,郝金凤,等.甜瓜β-氨基己糖苷酶基因家族的全基因组分析[J].广东农业科学,2014,41(8):185-188.
[4] GAO Y,WELLS L,COMER F I,et al. Dynamic Oglycosylation of nuclear and cytosolic proteins[J].Biol Chem,2001,276:9838-9845.
[5] LOVE D C,HANOVER J A. The hexosamine signaling pathway:deciphering the “O-GlcNAc code”[J].Sci Signaling,2005,312:13.
[6] SILVERSTONE A L,TSENG T,SWAIN SM,et al.Functional analysis of SPINDLY in gibberellin signaling in Arabidopsis[J].Plant Physiol,2007,143:987-1000.
[7] OLSZEWSKI N,SUN T P,GUBLER F.Gibberellin signaling biosynthesis,catabolism,&response pathways[J]. The Plant Cell Online,2002,14(S1):61-80.
[8]温都苏,阿拉塔,于斌.内蒙古野生黄花苜蓿种质资源及其开发利用前景[J].畜牧与饲料科学,2004,25(6):72-74.
[9]曹丽军.β-hex酶在沙红桃果实软化中作用的研究[D].杨凌:西北农林科技大学,2015.
[10]EVA L,CHRISTIANE V,MARTIN P,et al.β-NAcetylhexosaminidases HEXO1 and HEXO3 Are responsible for the formation of paucimannosidic Nglycans in Arabidopsis thaliana[J].The Journal of Biological Chemistryvol,2011,286:10793-10802.
[11]PRIEM B,GITTI R,BUSH C A,et al. Structure of ten free N-glycans in ripening tomato fruit(arabinose is a constituent of a plant N-glycan)[J].Plant Physiol,1993,102:445-458.
[12]JAGADEESH B H,PRABHA T N,SRINIVASAN K.Activities ofβ-hexosaminidase andα-mannosidase during development and ripening of bell capsicum(Capsicum annuum var. variata)[J].Plant Sci,2004,167:1263-1271.
[13]WANG J,ZHAO Y,RAY I,et al. Transcriptome responses in alfalfa associated with tolerance to intensive animal grazing[J]. Scientific Reports,2016,6(1):19438.