本文選題：川牛膝　切入點(diǎn)：ObgC　出處：《中國(guó)中藥雜志》2016年14期

【摘要】：根據(jù)川牛膝基因組數(shù)據(jù)提供的基因序列合成特異性引物,利用常規(guī)PCR方法和cDNA末端快速擴(kuò)增(RACE)技術(shù)克隆川牛膝Obg C(Gene Bank登錄號(hào)KU847910)全長(zhǎng)cDNA序列,克隆獲得2 226 bp全長(zhǎng)CoObgC序列,開(kāi)放閱讀框?yàn)? 818 bp,編碼605個(gè)氨基酸序列,預(yù)測(cè)CoObgC蛋白相對(duì)分子質(zhì)量為66.39 k Da,等電點(diǎn)p I 5.35,為穩(wěn)定蛋白,并進(jìn)行多重序列比對(duì)和構(gòu)建系統(tǒng)進(jìn)化樹(shù)分析。以川牛膝actin為內(nèi)參,采用實(shí)時(shí)熒光定量PCR(qRT-PCR)分析CoObgC基因在川牛膝根、莖、葉、花4種組織中表達(dá)特征,結(jié)果顯示在葉片中表達(dá)豐度最高,其次為根、花、莖;構(gòu)建p CABIA2300-CoObgC重組載體,利用農(nóng)桿菌介導(dǎo)法在煙草中進(jìn)行瞬時(shí)表達(dá),激光掃描共聚焦顯微鏡觀察顯示川牛膝CoObgC定位于葉綠體。該研究為進(jìn)一步解析Obg基因的結(jié)構(gòu)和功能,開(kāi)展川牛膝分子生物學(xué)研究奠定基礎(chǔ)。
[Abstract]:Specific primers were synthesized from Genomic data of Achyranthes bidentata. The full-length cDNA sequence of Obg C(Gene Bank (KU847910) was cloned by routine PCR method and rapid amplification of cDNA terminal cDNA. The full-length CoObgC sequence was obtained.The open reading frame is 1 818 BP, encoding 605 amino acid sequences. The predicted molecular weight of CoObgC protein is 66.39kDa. the isoelectric point I 5.35 is a stable protein, and the multiple sequence alignment and phylogenetic tree analysis are carried out.The expression characteristics of CoObgC gene in root, stem, leaf and flower of Achyranthes bidentata were analyzed by real-time fluorescence quantitative analysis with actin as internal reference. The results showed that the expression of CoObgC gene was the highest in the leaves, followed by the roots, flowers and stems, and the recombinant vector of p CABIA2300-CoObgC was constructed.Transient expression was carried out in tobacco by Agrobacterium tumefaciens. Laser scanning confocal microscopy showed that CoObgC was located in chloroplast of Achyranthes bidentata.This study laid a foundation for the further analysis of the structure and function of Obg gene and the development of molecular biology of Achyranthes bidentata.
【作者單位】： 四川農(nóng)業(yè)大學(xué)農(nóng)學(xué)院;四川農(nóng)業(yè)大學(xué)新農(nóng)村發(fā)展研究院;
【基金】：國(guó)家“十二五”科技支撐計(jì)劃項(xiàng)目(2011BAI13B02-7)
【分類號(hào)】：S567.239
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本文編號(hào)：1706517