【摘要】：在靈芝(Ganoderma lingzhi)中異源表達(dá)透明顫菌血紅蛋白(Vitreoscilla hemoglobin,VHb)基因,采用一氧化碳差異波譜分析的方法檢測轉(zhuǎn)基因菌株的生物活性,并對(duì)野生型菌株和工程菌株進(jìn)行發(fā)酵,通過熒光定量PCR對(duì)靈芝多糖生物合成途徑上的葡萄糖磷酸變位酶(α-phosphoglucomutase,PGM)、尿苷二磷酸葡萄糖焦磷酸化酶(uridinediphosphate glucose pyrophosphorylase,UGP)和β-1,3-葡聚糖合酶(β-1,3-glucan synthase,GLS)3個(gè)基因的轉(zhuǎn)錄水平進(jìn)行分析。研究表明:透明顫菌血紅蛋白基因(Vitreoscilla hemoglobin gene,vgb)在靈芝中能夠成功表達(dá),并且具有生物活性;工程菌株中胞外多糖的最高產(chǎn)量達(dá)0.83g/L,比野生型菌株提高了88.6%;與野生型菌株相比,工程菌株中多糖生物合成途徑上PGM、UGP和GLS基因的相對(duì)表達(dá)量分別是1.52、1.55和3.85。異源表達(dá)透明顫菌血紅蛋白基因是提高靈芝胞外多糖產(chǎn)量的一種有效方法。
[Abstract]:Vitreoscilla hemoglobin VHb gene was expressed in Ganoderma lingzhi. The biological activity of transgenic strain was detected by carbon monoxide differential spectrum analysis, and the wild type strain and engineering strain were fermented. The transcription levels of three genes, 偽 -phosphoglucomutase (PGM), uridine diphosphate glucose pyrophosphorylase (uridinediphosphate glucose) and 尾 -1n 3-glucan synthase (GLS), in the biosynthesis pathway of Ganoderma lucidum were analyzed by fluorescence quantitative polymerase chain reaction (FQ-PCR). The results showed that Vitreoscilla hemoglobin gene VGB could be successfully expressed in Ganoderma lucidum and had biological activity, the highest yield of exopolysaccharide in engineering strain was 0.83g / L, which was 88.6g / L higher than that of wild-type strain. The relative expression levels of PGMU UGP and GLS genes in the polysaccharide biosynthesis pathway were 1.521.55 and 3.85, respectively. Heterologous expression of fibrillation hemoglobin gene is an effective method to increase the production of Ganoderma lucidum extracellular polysaccharides.
【作者單位】： 昆明理工大學(xué)生命科學(xué)與技術(shù)學(xué)院;昆明理工大學(xué)理學(xué)院;
【基金】：國家自然科學(xué)基金項(xiàng)目(31360495和21566016)資助
【分類號(hào)】：S567.31
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本文編號(hào)：2123728