Inducible EGFP expression under the control of the nitrate reductase gene promoter in transgenic Dunaliella salina
作者： Jie Li , Lexun Xue , Hongxia Yan , Hongtao Liu , Jianyang Liang
DOI: 10.1007/S10811-007-9197-0
关键词: Biology 、 Heterologous expression 、 Molecular biology 、 Heterologous 、 Nitrate reductase 、 Promoter 、 Gene 、 Dunaliella salina 、 Expression cassette 、 Transgene
摘要: The upstream region of the nitrate reductase (NR) gene is able to provide an ideal promoter for inducible expression heterologous genes because NR transcripts are induced by and repressed ammonium. To improve proteins in transgenic Dunaliella salina, we developed a novel vector with ability switch on or off using gene. In present study, 1.2-kb fragment containing (Pnr) 0.9-kb terminator (Tnr) were cloned from genome D. salina. results mobility shift assays indicated that GATA element isolated Pnr was involved nitrogen-dependent regulation. Additionally, transcription start site salina located at 133 bp codon ATG. Gene splicing overlap extension (SOEing) employed construct recombinant vector, p7NET, Pnr-EGFP-Tnr cassette. On day 2 after p7NET introduced into cells, EGFP-expressing transformants appeared, which EGFP but RT-PCR PCR-Southern blots showed correctly transcribed findings study suggest may be used drive
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