本文選題：青稞　切入點(diǎn)：類(lèi)黃酮O-甲基轉(zhuǎn)移酶　出處：《核農(nóng)學(xué)報(bào)》2017年12期

【摘要】：類(lèi)黃酮O-甲基轉(zhuǎn)移酶(FOMT)是一種在植物甲基化黃酮代謝合成中有著重要作用的酶,但在中國(guó)青藏高原的特色作物青稞中研究較少。為了更好了解該酶及編碼基因在青稞中的生理功能及作用,以高總黃酮青稞品系94-19-1為材料,通過(guò)RT-PCR擴(kuò)增、克隆得到青稞OMT1基因的ORF序列。結(jié)果表明,該基因ORF長(zhǎng)1 071 bp,編碼356個(gè)氨基酸,預(yù)測(cè)蛋白分子量為38.65 KDa,等電點(diǎn)為5.33。序列比對(duì)分析發(fā)現(xiàn),所克隆的基因與NCBI數(shù)據(jù)庫(kù)收錄的大麥Hv OMT1基因有99.44%的一致性,氨基酸序列存在5個(gè)殘基差異,編碼的蛋白序列具有OMT蛋白家族典型的保守結(jié)構(gòu)域。進(jìn)一步將該基因連接到原核表達(dá)載體轉(zhuǎn)化大腸桿菌,最終成功將該基因所編碼的蛋白在大腸桿菌誘導(dǎo)表達(dá),并利用親和層析柱分離技術(shù),將該表達(dá)蛋白進(jìn)行了純化。這為進(jìn)一步研究該酶蛋白功能和選育高品質(zhì)青稞品種奠定了基礎(chǔ)。
[Abstract]:Flavonoid O-methyltransferase (FOMT) is an enzyme that plays an important role in the metabolism and synthesis of plant methylated flavonoids. In order to better understand the physiological function and function of the enzyme and coding gene in highland barley, the high flavonoid highland barley line 94-19-1 was used as material and amplified by RT-PCR. The ORF sequence of OMT1 gene of highland barley was cloned. The results showed that the ORF length of the gene was 1071 BP, encoding 356 amino acids, the molecular weight of predicted protein was 38.65 KDaand the isoelectric point was 5.33. The cloned gene was 99.44% consistent with that of barley Hv OMT1 gene collected in NCBI database, and the amino acid sequence was different from that of barley Hv OMT1 gene. The encoded protein sequence has the typical conserved domain of OMT protein family. The gene was further ligated into prokaryotic expression vector and transformed into Escherichia coli. Finally, the protein encoded by the gene was successfully induced to express in E. coli. The expressed protein was purified by affinity chromatography, which laid a foundation for further study on the function of the enzyme and breeding of highland barley varieties with high quality.
【作者單位】： 四川農(nóng)業(yè)大學(xué)農(nóng)學(xué)院/作物遺傳育種學(xué)系大麥青稞研究中心;
【基金】：國(guó)家現(xiàn)代農(nóng)業(yè)產(chǎn)業(yè)技術(shù)體系(大麥青稞體系)建設(shè)專(zhuān)項(xiàng)基金(CARS-05)
【分類(lèi)號(hào)】：Q943.2;S512.3
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本文編號(hào)：1659819