水稻塑性葡萄糖-6-磷酸脱氢酶基因的分离与表达分析

HOU Fu-Yun , HUANG Ji , LU Ju-Fei , WANG Zhou-Fei , ZHANG Hong-Sheng
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引用次数: 8

摘要

葡萄糖-6-磷酸脱氢酶是一种戊糖磷酸途径的限速酶,存在于高等植物的胞质和质体中。从水稻(Oryza sativa L.)中分离到一个编码塑性葡萄糖-6-磷酸脱氢酶的新基因,命名为OsG6PDH2。通过半定量RT-PCR方法发现,OsG6PDH2 mRNA在水稻叶片、茎、未成熟穗和花穗中表达较弱,在根系中表达稍高。而暗处理能显著诱导OsG6PDH2在水稻幼苗中的表达。将OsG6PDH2的完整开放阅读框(ORF)插入pET30a(+)中,在大肠杆菌BL21 (DE3)中表达。转化细菌细胞的酶活性测定表明,OsG6PDH2编码产物具有典型的葡萄糖-6-磷酸脱氢酶功能。
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Isolation and Expression Analysis of Plastidic Glucose-6-phosphate Dehydrogenase Gene from Rice (Oryza sativa L.)

Glucose-6-phosphate dehydrogenase is a rate-limiting enzyme of pentose phosphate pathway, existing in cytosolic and plastidic compartments of higher plants. A novel gene encoding plastidic glucose-6-phosphate dehydrogenase was isolated from rice (Oryza sativa L.) and designated OsG6PDH2 in this article. Through semiquantitative RT-PCR approach it was found that OsG6PDH2 mRNA was weakly expressed in rice leaves, stems, immature spikes or flowered spikes, and a little higher in roots. However, the expression of OsG6PDH2 in rice seedlings was significantly induced by dark treatment. The complete opening reading frame (ORF) of OsG6PDH2 was inserted into pET30a (+), and expressed in Escherichia coli strain BL21 (DE3). The enzyme activity assay of transformed bacterial cells indicated that OsG6PDH2 encoding product had a typical function of glucose-6-phosphate dehydrogenase.

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