The human cytomegalovirus immediate early gene promoter is a strong promoter in cultured Drosophila melanogaster cells.
The human cytomegalovirus immediate early gene promoter is a strong promoter in cultured Drosophila melanogaster cells.
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DOI：
10.1093/nar/15.5.2392
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发表时间：
1987-03
影响因子：
14.9
通讯作者：
J. Sinclair
中科院分区：
文献类型：
--
作者：
J. Sinclair
Analysis of promoters capable of directing expression of the chlo ical acetyl transferase (CAT) gene in cultured Drosophila cells has thmn that the huf n cytcaslovirus major immediate early (HCI IE) gene promter/enhncer region is a very strong prcwter in the Drosophila Sc cell line. Previously heterologous praiters fran other vertebrate DNA and RNiA tsour viruses have been shown to function poorly in D phila cells (1,2). The HOW IE enhancer is one of the strongest eukaryotic enhancers and drives high levels of expression of eukaryotic genes (3). plEPiCat containing the HOUK prcuter/enhancer unit (G. Wilkinson and A. Akrigg, uwpblitied) is expressed in D lia c cells as efficiently as pCV2Cat, which, in our hands, is the most powerful CAT expression vector for Dros~hila (4). It is interesting to note that pIEPlCat is not expressed in a D. hydei cell line (DH33) even though this cell line is capable of very high levels of expression of pCV2Cat, suesting that Drosophila specific factors may be regulating the HQV IE pranoter differentially in Kc and 1D33 cells. A good heterologous expression vector will be extremely useful as a control, reference gene in transient expression assays analysing Dro pila regulatory sequences and sliafld also provide the basis for a high frequency transforming daninant selectable marker. Figure Legend a)pIEPlCat; the Cat gene (hatched box) under tie control of 302bp of 5' upstream sequences of the HCK IE gene and 295bp of 3' downstream sequences. b)After transfection (4) with pIEPlCat (a) or pCV2Cat (b) Sc cells (A) or 1 cell (B) were assayed for CAT acvitity (3). Con are control untransfected cells. Conversion of chloraqenicol to acetylated products are shown as a a ** percentage of that observed for C2Cat in Kc cells (lOnmol of [wr chloranphenicol/90min/2x106 cells) and are; pCV2Cat in DH33 cells, 85% pIEPlCat in Kc cells, 84% and pIEPICat in D33 celTls , 2%. Acknowlegements: I gratefully acknowledge the gift of pIEPlCat fram Gavin Wilklino and lan Akrigg (PEL, Porton Down). Rleferences 1)DF Nolea, P.P. and Dlvid, I.B. (1983). Proc. Natl. Acad. Sci.USA, 86 7095-7098. 2) Burke, J.F.,et al(1984). 0 J.3 2549-2554. 3) Boshart, M. et al(1985). Cell 41 521-530. 4) Sinclair,J.H.,eC al(1986). E1W. 5 2349-2354.
