本文選題：三葉青　切入點：擴展蛋白基因　出處：《中草藥》2016年05期 　論文類型：期刊論文

【摘要】：目的克隆三葉青Tetrastigma hemsleyanum的擴展蛋白家族基因,對其進行器官表達分析,并初步探討其功能。方法根據(jù)Gen Bank中登錄的擴展蛋白基因保守區(qū)域序列設計1對簡并引物,以三葉青球形塊根c DNA為模板進行RT-PCR擴增,再結(jié)合RACE技術,獲得三葉青擴展蛋白基因全長c DNA序列,命名為Th-exp。采用半定量RT-PCR和熒光定量PCR分析Th-exp基因在不同器官中的表達情況。結(jié)果克隆獲得擴展蛋白家族基因,共782 bp,其中包含630 bp開放閱讀框,編碼209個氨基酸(Gen Bank登陸號KP693606)。Th-exp編碼的蛋白質(zhì)具有典型的擴展蛋白結(jié)構(gòu)特征,N端含有8個半胱氨酸的結(jié)構(gòu)域,C端含有4個保守的色氨酸結(jié)構(gòu)域,中間含有組氨酸功能域。Blast分析顯示,Th-exp與葡萄、醉蝶花中擴展蛋白基因具有相似性。半定量RT-PCR和熒光定量PCR分析表明,Th-exp在三葉青葉、莖、普通細根和球形塊根中均有表達,但球形塊根中表達強于其他器官。結(jié)論克隆得到三葉青擴展蛋白家族基因Th-exp全長c DNA,初步預測其參與了三葉青塊根的發(fā)育過程。
[Abstract]:Objective to clone the extended protein family genes of three Ye Qing Tetrastigma hemsleyanum, analyze their organ expression, and explore their functions. Methods A pair of degenerate primers were designed according to the conserved region of the extension protein gene entered in Gen Bank. The full-length c DNA sequence of the c DNA gene of three Ye Qing globular tubers was obtained by RT-PCR amplification with RACE technique. The expression of Th-exp gene in different organs was analyzed by semi-quantitative RT-PCR and fluorescence quantitative PCR. The protein encoded by the KP693606).Th-exp encoding 209 amino acid sequence Gen Bank has typical structural characteristics of extended protein. The N-terminal domain containing 8 cysteine has four conserved tryptophan domains at the C-terminal. The intermediate histidine domain. Blast analysis showed that Th-exp gene was similar to that of grape. Semi-quantitative RT-PCR and fluorescence quantitative PCR analysis showed that Th-exp was expressed in green leaves, stems, fine roots and globular tubers. Conclusion the full-length Th-exp cDNA of the three Ye Qing extended protein family gene was obtained and it was preliminarily predicted that it was involved in the development of the root block of the three Ye Qing tubers.
【作者單位】： 杭州師范大學生命與環(huán)境科學學院;
【基金】：浙江省藥用植物種質(zhì)改良與質(zhì)量控制技術重點實驗室開放項目(201302)
【分類號】：S567.2;Q943.2

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本文編號：1647005