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phbB、phbC基因克隆、序列分析及植物表达载体的构建 被引量:21
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作者 谢安勇 崔晓江 宋艳茹 《Acta Botanica Sinica》 CSCD 1995年第8期581-588,共8页
利用聚合酶链式反应(PCR)技术,从真养产碱杆菌Alcaligenes eutrophus H16 染色体DNA 中扩增并克隆了调控聚-β-羟基丁酸(poly-β-hydroxybutyrate, PHB)生物合成的两个... 利用聚合酶链式反应(PCR)技术,从真养产碱杆菌Alcaligenes eutrophus H16 染色体DNA 中扩增并克隆了调控聚-β-羟基丁酸(poly-β-hydroxybutyrate, PHB)生物合成的两个关键酶基因:依赖NADPH 的乙酰乙酰CoA 还原酶基因(phbB)和PHB合成酶基因(phbC)。限制性内切酶图谱和核苷酸序列分析证实了克隆结果,并表明所克隆的基因与国外报道的有很高的同源性。经过基因拼接,构建了块茎特异性表达的高等植物表达载体pPSAGB(嵌合phbB)、pBIBGC(嵌合phbC)和pPSAGCB(嵌合phbB和phbC双基因) 展开更多
关键词 聚β羧基丁酸 PHBB phbc 基因克隆 植物表达载体
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Obtainment of Transgenic Tobacco Harboring phbA , phbB and phbC Genes by Twice Transformation 被引量:4
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作者 张景昱 叶梁 +1 位作者 李丽 宋艳茹 《Acta Botanica Sinica》 CSCD 2001年第1期59-62,共4页
To avoid the long time required for conventional sexual crossing, transgenic tobacco (Nicotiana tabacum L.) plants harboring phbA gene (encoding 3_ketothiolase) were used as the target plant for the second transfo... To avoid the long time required for conventional sexual crossing, transgenic tobacco (Nicotiana tabacum L.) plants harboring phbA gene (encoding 3_ketothiolase) were used as the target plant for the second transformation mediated by Agrobacterium tumefaciens (Smith et Townsend) Conn LBA4404 containing pZCB which was constructed by linking phbB (encoding acetoacetyl_CoA reductase), phbC (encoding PHB synthase) and ctp sequence to pBIB_HYG under the control of CaMV 35S promoter. The hygromycin resistant transformants were morphologically normal and stable integration of phbB and phbC was confirmed by PCR and PCR_Southern. Moreover, RT_PCR_DNA hybridization analysis showed that 6.67% of the transformed tobacco plants could express both phbB and phbC at transcriptional level. 展开更多
关键词 poly (3_hydroxybutyrate) (PHB) twice transformation transgenic tobacco
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黄淮海平原典型潮土的酸碱缓冲性能 被引量:30
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作者 黄平 张佳宝 +1 位作者 朱安宁 张丛志 《中国农业科学》 CAS CSCD 北大核心 2009年第7期2392-2396,共5页
【目的】通过对土壤酸碱缓冲容量的研究,可以评价土壤抵抗酸化和碱化的能力,也可以对土壤酸化过程进行预测和调控。【方法】采用酸碱滴定法获取土壤酸碱滴定曲线,并通过分段拟合计算土壤酸碱缓冲容量。【结果】酸滴定曲线和碱滴定曲线... 【目的】通过对土壤酸碱缓冲容量的研究,可以评价土壤抵抗酸化和碱化的能力,也可以对土壤酸化过程进行预测和调控。【方法】采用酸碱滴定法获取土壤酸碱滴定曲线,并通过分段拟合计算土壤酸碱缓冲容量。【结果】酸滴定曲线和碱滴定曲线在整体上并不呈线性,但分段的酸和碱滴定曲线的线性相关系数均大于0.97。通过分段拟合得出土壤酸、碱缓冲容量分别为158.71和25.02mmol·kg-1。【结论】酸碱滴定法适用于黄淮海平原典型潮土酸碱缓冲容量测定。根据缓冲体系的划分和土壤基本性质可以看出,碳酸盐是最重要的酸缓冲体系,而土壤有机质贡献相对较小。 展开更多
关键词 典型潮土 黄淮海平原 pH缓冲容量
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Acid and Alkali Buffer Capacity of Typical Fluvor-Aquic Soil in Huang-Huai-Hai Plain 被引量:11
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作者 HUANG Ping ZHANG Jia-bao +1 位作者 ZHU An-ning ZHANG Cong-zhi 《Agricultural Sciences in China》 CSCD 2009年第11期1378-1383,共6页
Soil acid and alkali buffer capacity, as a major indicator for evaluating its vulnerability and resistibility to acidification and alkalization, is an important factor affecting the sustainable agriculture, through kn... Soil acid and alkali buffer capacity, as a major indicator for evaluating its vulnerability and resistibility to acidification and alkalization, is an important factor affecting the sustainable agriculture, through knowledge on which soil acidification process can be predicted and modified. In this study, titration curve method was adopted to investigate the pH buffer capacity (pHBC) of fluvor-aquic soil, and separate titration curves were established by adding incremental amounts of either standardized hydrochloric acid (HC1) (0.12 mol L^-1) or sodium hydroxide (NaOH) (0.10 mol L^-1) to soil suspended in deionized water (soil:solution = 1:5). Soil pH was measured after 7 d resuspension and isothermal equilibrium (T = 25℃). Linear regressions were fitted to the linear portion of each titration curve and the slopes of these lines were derived as the soil pHBC. The results showed that significant correlations between the amounts of adding acid or alkali and each pH change were presented, and titration curve method was feasible for measurement of pHBC on typical fluvor-aquic soil in Huang-Huai-Hai Plain, and the coefficients of determination were higher than the similar researches on acid soil (R^2 = 0.96). The slope-derived pHBC of acid and alkali were 158.71 and 25.02 mmol kg^-1, respectively. According to the classification of soil buffer systems, the soil tested belongs to the calcium carbonate buffer system, carbonates contribute the most to pHBC, and the contribution of soil organic matter relatively less than it. 展开更多
关键词 fluvor-aquic soil Huang-Huai-Hai Plain pH buffer capacity phbc
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Hsp70A-RBCS2融合启动子调控PHB合成酶基因在莱茵衣藻中的表达 被引量:1
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作者 王潮岗 胡章立 《中国科学(C辑)》 CSCD 北大核心 2009年第8期775-782,共8页
利用Hsp70A-RBCS2融合启动子构建了新型的莱茵衣藻表达载体,并获得了聚-β-羟基丁酸(PHB)合成酶基因(phbC)的衣藻表达载体p105C124和pH105C124.通过"珠磨法"分别将上述两个衣藻表达载体导入细胞壁缺陷的莱茵衣藻CC-849(Chlamy... 利用Hsp70A-RBCS2融合启动子构建了新型的莱茵衣藻表达载体,并获得了聚-β-羟基丁酸(PHB)合成酶基因(phbC)的衣藻表达载体p105C124和pH105C124.通过"珠磨法"分别将上述两个衣藻表达载体导入细胞壁缺陷的莱茵衣藻CC-849(Chlamydomonas reinhardtii CC-849)中,得到了具有Zeomycin抗性的转基因藻株.Hsp70A-RBCS2启动子介导的外源基因遗传转化效率明显高于RBCS2启动子,Southern杂交结果显示,phbC基因以低拷贝数整合进莱茵衣藻的基因组DNA中.在光照下,Hsp70A-RBCS2融合启动子能够有效调控phbC基因在莱茵衣藻中的转录和翻译,得到的蛋白产物具有PHB合成酶活性,40℃热激诱导可使PHB合成酶的酶活性提高到1.8倍.因此,Hsp70A-RBCS2融合启动子可使phbC基因在莱茵衣藻中实现可诱导的表达,该研究对利用衣藻合成PHB具有重要的学术意义,进一步的研究将通过"共转化"法获得二价或三价的转基因藻,最终实现在转基因藻中合成PHB. 展开更多
关键词 转基因衣藻 phbc基因 Hsp70A-RBCS2启动子 聚-β-羟基丁酸(PHB)
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利用转基因衣藻合成聚-β-羟基丁酸的研究 被引量:1
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作者 王潮岗 胡章立 《深圳特区科技》 2005年第11期211-216,共6页
聚-β-羟基丁酸(polyhydroxybutyric acid,PHB)是发现最早且研究最透彻的一种生物可降解塑料。莱茵衣藻(Chlamydomonas reinhardtii)素有“光合酵母”之称,是理想的转基因受体生物。通过转基因技术将PHB 生物合成的关键酶基因导入莱茵衣... 聚-β-羟基丁酸(polyhydroxybutyric acid,PHB)是发现最早且研究最透彻的一种生物可降解塑料。莱茵衣藻(Chlamydomonas reinhardtii)素有“光合酵母”之称,是理想的转基因受体生物。通过转基因技术将PHB 生物合成的关键酶基因导入莱茵衣藻,利用光合作用合成 PHB,降低 PHB 的生产成本。从真养产碱杆菌(Alcaligenes eutrophus)分离出 phbB 和 phbC 基因,然后构建 phbB 和 phbC 基因的衣藻表达载体 p105B124和 pH105C124。通过“珠磨法”遗传转化技术共转化 p105B124和 pH105C124,获得了二价转基因衣藻。分子检测结果表明 phbB 和 phbC 基因均已整合到莱茵衣藻基因组中。随后进行结晶紫染色和显微镜观察转基因藻,发现二价转化子的核区和细胞膜附近分布有白色空泡;进一步的电子显微镜观察结果表明白色空泡是由细胞中合成的 PHB 聚集而成,电镜下观察到由 PHB 形成的明亮的圆形颗粒。光照(90μE/m^2/s)条件下培养转基因藻,出现生长受抑制现象,这可能是由于 PHB 颗粒在藻细胞内随机分布,干扰了细胞正常的生命活动。 展开更多
关键词 转基因藻 phbB基因 phbc基因 聚-β-羟基丁酸(PHB)
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