Candida glabrata shuttle vectors suitable for translational fusions to lacZ and use of β-galactosidase as a reporter of gene expression
Candida glabrata shuttle vectors suitable for translational fusions to lacZ and use of β-galactosidase as a reporter of gene expression
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DOI：
10.1016/s0378-1119(00)00065-2
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发表时间：
2000-04-04
期刊：
影响因子：
3.5
通讯作者：
Mühlschlegel, FA
中科院分区：
文献类型：
--
作者：
El Barkani, A;Haynes, K;Mühlschlegel, FA
研究了由大肠杆菌 lacZ 基因编码的 β-半乳糖苷酶作为光滑 C. glabrata 基因表达报告基因的功能。光滑 C. 光滑/E。构建了大肠杆菌穿梭载体，其中含有光滑念珠菌 CEN-ARS 盒，以允许质粒的规则分离和附加型复制，以及大肠杆菌的 lacZ 编码序列。通过将启动子和来自光滑 C. glabrata 的 HIS3 基因的 5' 编码区定向插入 lacZ 基因上游，验证了 C. glabrata 中 β-半乳糖苷酶的功能。通过将铜控制的 MTII 基因的启动子与 lacZ 报告基因融合，我们发现可以在光滑 C. glabrata 中差异诱导 P-半乳糖苷酶活性。通过间接过滤测定定性检测β-半乳糖苷酶报告活性，并从透化细胞中定量检测。 (C) 2000 Elsevier Science B.V. 保留所有权利。
The functionality of P-galactosidase encoded by the E. coli lacZ gene as a reporter of gene expression in C. glabrata was investigated. C. glabrata/E. coli shuttle vectors were constructed, containing both a C. glabrata CEN-ARS cassette, to allow regular segregation and episomal replication of the plasmids, and the lacZ coding sequence of E, coli. The functionality of P-galactosidase in C. glabrata was verified by inserting the promoter and the 5' coding region of the HIS3 gene from C. glabrata directionally upstream of the lacZ gene. By fusing the promoter of the copper-controlled MTII gene to the lacZ reporter, we showed that P-galactosidase activity can be differentially induced in C. glabrata. P-galactosidase reporter activities were detected qualitatively by an indirect filter assay and quantitatively from permeabilized cells. (C) 2000 Elsevier Science B.V. All rights reserved.