Ross S R, Hsu C L, Choi Y, Mok E, Dudley J P
Department of Biochemistry, University of Illinois, Chicago 60612.
Mol Cell Biol. 1990 Nov;10(11):5822-9. doi: 10.1128/mcb.10.11.5822-5829.1990.
Mouse mammary tumor virus (MMTV) is an endogenous murine retrovirus that is expressed in the epithelial cells of the mammary and salivary glands, lungs, kidneys, and seminal vesicles and in the lymphoid cells of the spleen and thymus. Several studies have shown that the long terminal repeat (LTR) of this virus can direct the expression of reporter genes to the same tissues in transgenic mice. To determine whether multiple regulatory elements within the LTR are involved in this tissue-specific expression, we have established lines of transgenic mice containing transgenes that have deletions in the MMTV LTR. Deletions of all LTR sequences upstream of -364 or of LTR sequences from -165 to -665 both result in the expression of linked reporter genes such as the simian virus 40 early region or the bacterial enzyme chloramphenicol acetyltransferase in novel sites, such as the heart, brain, and skeletal muscle; expression of endogenous MMTV and transgenes containing the full-length LTR is not detected in these organs. Negative regulation appears to involve more than one region, since deletion of sequences between either -201 and -471 or -201 and -344, as well as sequences upstream of -364, results in inappropriate expression in heart, brain, and skeletal muscle. Therefore, a negative regulatory element(s) in the MMTV LTR can suppress transcription from the viral promoter in several different organs. This represents the first example of generalized negative regulatory elements that act in many different tissues in transgenic mice to prevent inappropriate expression of a gene.
小鼠乳腺肿瘤病毒(MMTV)是一种内源性鼠逆转录病毒,在乳腺、唾液腺、肺、肾和精囊的上皮细胞以及脾脏和胸腺的淋巴细胞中表达。多项研究表明,该病毒的长末端重复序列(LTR)可将报告基因的表达导向转基因小鼠的相同组织。为了确定LTR内的多个调控元件是否参与这种组织特异性表达,我们建立了含有在MMTV LTR中具有缺失的转基因的转基因小鼠品系。-364上游所有LTR序列的缺失或-165至-665的LTR序列的缺失均导致连接的报告基因如猿猴病毒40早期区域或细菌酶氯霉素乙酰转移酶在新的位点如心脏、大脑和骨骼肌中表达;在这些器官中未检测到内源性MMTV和含有全长LTR的转基因的表达。负调控似乎涉及多个区域,因为-201至-471或-201至-344之间的序列以及-364上游的序列的缺失导致在心脏、大脑和骨骼肌中出现不适当的表达。因此,MMTV LTR中的一个或多个负调控元件可抑制病毒启动子在几个不同器官中的转录。这代表了在转基因小鼠的许多不同组织中起作用以防止基因不适当表达的广义负调控元件的第一个例子。