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转基因蔓越莓中可检测到的β-葡萄糖醛酸酶活性受内源性抑制剂和植物发育的影响。

Detectable ,-glucuronidase activity in transgenic cranberry is affected by endogenous inhibitors and plant development.

作者信息

Serres R, McCown B, Zeldin E

机构信息

Department of Horticulture, University of Wisconsin-Madison, Madison, WI 53706, USA Fax no: +1-608-262-4743 E-mail:

出版信息

Plant Cell Rep. 1997 Jun;16(9):641-646. doi: 10.1007/s002990050294.

Abstract

Extracts of cranberry, rich in flavonols and proanthocyanins, inactivated β-glucuronidase (GUS) in assays using either purified bacterial GUS or preparations of transgenic tobacco (Nicotiana tabacum L.) or transgenic cranberry (Vaccinium macrocarpon Ait.) expressing the gusA gene. Histochemical GUS assays produced random and generally unpredictable staining. The addition of polyvinylpolypyrrolidone (PVPP) during the preparation of transgenic cranberry leaf extracts increased the detectable GUS activity in fluorogenic assays more than 200-fold. Detectable GUS activity varied among transclones and also within a transclone depending on the developmental and physiological state of the tissue, as well as the growth environment. Use of PVPP altered the relative ranking of plants based on their total transgenic enzyme activity and resulted in different conclusions as to the effects of genotype or growth environment on transgene expression.

摘要

富含黄酮醇和原花青素的蔓越莓提取物,在使用纯化的细菌β-葡萄糖醛酸酶(GUS)或表达gusA基因的转基因烟草(Nicotiana tabacum L.)或转基因蔓越莓(Vaccinium macrocarpon Ait.)制剂的试验中,可使β-葡萄糖醛酸酶(GUS)失活。组织化学GUS分析产生了随机且通常不可预测的染色。在制备转基因蔓越莓叶提取物过程中添加聚乙烯聚吡咯烷酮(PVPP),可使荧光分析中可检测到的GUS活性提高200多倍。可检测到的GUS活性在不同转基因克隆之间存在差异,并且在一个转基因克隆内也因组织的发育和生理状态以及生长环境而异。使用PVPP改变了基于植物总转基因酶活性的相对排名,并导致关于基因型或生长环境对转基因表达影响的不同结论。

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