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铁皮石斛均一化全长cDNA文库的构建与序列分析

Keywords: 铁皮石斛,均一化,cDNA文库,序列分析

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Abstract:

目的:为了获得药用植物铁皮石斛功能基因的信息和筛选功能基因。方法:以SMART(switchingmechanismat5'endofRNAtranscript)方式合成全长cDNA,结合DSN(duplex-specificnuclease)均一化技术,构建铁皮石斛茎组织的均一化全长cDNA文库。结果:该文库重组率为93.9%,文库滴度1.3×106cfu·mL-1,插入片段平均大小1.5kb左右。随机挑取163个阳性克隆进行单侧测序,通过生物信息学分析,获得150个单一序列,其中147个序列与相应的同源蛋白匹配,GO(geneontology)分类表明这些序列参与各种生化途径。8个序列包含了完整编码框,可判断为全长基因。结论:成功构建了铁皮石斛茎组织均一化全长cDNA文库,为满足铁皮石斛代谢途径等功能基因筛选和基因信息研究奠定了基础。

References

[1]  蔡刘体,胡重怡,张永春,等.烤烟品种南江3 号均-化全长cDNA 文库构建[J].生物技术,2011,21(2):45.
[2]  Ke T,Dong C H,Mao H,et al. Construction of a normalized full-length cDNA library of sesame dseveloping seed by DSN and SMARTTM[J]. Agr Sci Chin,2011,10(7): 1004.
[3]  曾日中,段翠芳,聂智毅,等. 橡胶树胶乳均一化全长cDNA 文库的构建与EST 测序分析[J]. 中国农业科学,2011,44(4):683.
[4]  陈士林,孙永珍,徐江,等. 本草基因组计划研究策略[J]. 药学学报,2010,45(7): 807.
[5]  Mohd-Hairul A R,Namasivayam P,Abdullah J O,et al. Screening, isolation,and molecular characterization of putative fragrance-related transcripts from vanda mimi palmer[J]. Acta Physiol Plant,2011,33:1651.
[6]  Wang P,Heitman J. The cyclophilins[J]. Genome Biol,2005,6(7):226.
[7]  鲍永美,王州飞,张红生.植物SNARE蛋白的结构与功能[J].植物学通报,2005,22(6): 715.
[8]  Chen Z H,Sun X F,Tang K X. Cloning and expression of a novel cDNA encoding a mannose-binding lectin from Dendrobium officinale[J]. Toxicon,2005,45 :535.
[9]  孟衡玲,段承俐,萧凤回,等.铁皮石斛蔗糖合成酶基因的克隆及表达分析[J].中国中药杂志,2011,36(7):833.
[10]  Zhao P, Wang W J, Sun M X. Characterization and expression pattern analysis of DcNAC gene in somatic embryos of Dendrobium candidum Wall Ex Lindl[J]. Plant Cell, Tissue Organ Cult,2011,107:151.
[11]  Yang S H, Yu H, Goh, C J. Functional characterisation of a cytokinin oxidase gene DSCKX1 in dendrobium orchid[J]. Plant Mol Biol,2003, 51: 237.
[12]  Sattayasai N, Sudmoon R, Nuchadomrong S, et al. Dendrobium findleyanum agglutinin: production, localization, anti-fungal activity and gene characterization[J]. Plant Cell Rep,2009,28:1243.
[13]  Soares M B,Bonaldo M F,Jelene P,et al. Construction and characterization of a normalized cDNA library[J]. Proc Natl Acad Sci USA,1994,91(20):9228.
[14]  Shagin D A, Rebrikov D V, Kozhemyako V B, et al. A novel method for SNP detection using a new duplex-specific nuclease from crab hepatopancreas[J]. Genome Res, 2002,12:1935.
[15]  Zhulidov P A,Bogdanova E A,Shcheglov A S,et al. Simple cDNA normalization using kamchatka crab duplex-specific nuclease[J]. Nucl Acid Res,2004,32:e37.
[16]  Zhang Z Q,Xiao G,Tan T L,et al. Advances in normalized cDNA library and its applications[J]. Agr Sci Technol,2010,11 (4) :1.
[17]  包雪声,顺庆生,张申洪,等. 中国药用石斛图志[M]. 上海:科学技术文献出版社,2005.
[18]  李桂锋,李进进,许继勇,等. 铁皮石斛研究综述[J]. 中药材,2010,33(1):150.
[19]  魏凤娟.铁皮石斛组织培养与栽培技术研究进展[J].广东农业科学,2010,4:81.
[20]  朱平,王伟,程克棣. 药用植物功能基因[J]. 中国生物工程杂志,2004,24(2):3.
[21]  Seki M,Narusaka M,Kamiya A,et al. Function annotation of a full-length Arabidopsis cDNA collection[J]. Science,2002,296:11.
[22]  Zhu Y Y,Machleder E M,Chenchik A,et al. Reverse transcriptase template switching: a SMART approach for full-length cDNA library construction[J]. Biotechniques,2001,30,892.
[23]  Wellenreuther R,Schupp I,Poustka A,et al. German cDNA Consortium.SMART amplification combined with cDNA size fractionation in order to obtain large full-length clones[J]. BMC Genomics,2004,5:36.
[24]  王卓,殷晓敏,王家保,等. 香蕉根系均一化全长cDNA 文库的构建和鉴定[J]. 园艺学报,2011,38(2):91667.

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