EXPRESSION OF THE MURINE ALPHA-B-CRYSTALLIN GENE IS NOT RESTRICTED TO THE LENS
EXPRESSION OF THE MURINE ALPHA-B-CRYSTALLIN GENE IS NOT RESTRICTED TO THE LENS
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DOI：
10.1128/mcb.9.3.1083
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发表时间：
1989-03-01
影响因子：
5.3
通讯作者：
PIATIGORSKY, J
中科院分区：
文献类型：
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作者：
DUBIN, RA;WAWROUSEK, EF;PIATIGORSKY, J
克隆鼠α B-晶状体蛋白基因并检测其表达。在小鼠中，不仅在透镜中，而且在心脏、骨骼肌、肾和肺中检测到显著水平的α B-晶状体蛋白RNA：在脑和脾中分别检测到低水平和痕量水平。肺、脑和脾中的RNA种类比其他组织中的大400至500个碱基。在透镜、心脏、骨骼肌、肾脏和脑中的转录起始于相同的位置。构建小鼠α B-晶状体蛋白微型基因并将其导入小鼠的生殖系，并且证明其表达与内源基因的表达平行。在透镜、心脏和骨骼肌中总是检测到转基因RNA，而在肾和肺中的表达是可变的;仍不确定脑和脾中是否存在转基因表达。这些结果表明，控制α B-晶状体蛋白基因表达的调节序列位于转录起始位点上游666个碱基对的序列和poly（A）添加位点下游2.4个碱基对的序列之间，并且不位于内含子内。用一系列α B-晶状体蛋白小基因缺失突变体进行的转染研究揭示，在原代鸡胚透镜细胞中有效表达需要位置-222和-167之间的序列;在该体外系统中表达需要poly（A）加成信号下游的序列。
The murine .alpha.B-crystallin gene was cloned and its expression was examined. In the mouse, significant levels of .alpha.B-crystallin RNA were detected not only in lens but also in heart, skeletal muscle, kidney, and lung: low and trace levels were detected in brain and spleen, respectively. The RNA species in lung, brain, and spleen was 400 to 500 bases larger than that in the other tissues. Transcription in lens, heart, skeletal muscle, kidney, and brain initiated at the same position. A mouse .alpha.B-crystallin mini-gene was constructed and was introduced into the germ line of mice, and its expression was demonstrated to parallel that of the endogenous gene. Transgene RNA was always detected in lens, heart, and skeletal muscle, while expression in kidney and lung was variable; it remains uncertain whether there is transgene expression in brain and spleen. These results demonstrate that regulatory sequences controlling expression of the .alpha.B-crystallin gene lie between sequences 666 base pairs upstream of the transcription initiation site and 2.4 kilobase pairs downstream of the poly(A) addition site and are not located within the introns. Transfection studies with a series of .alpha.B-crystallin mini-gene deletion mutants revealed that sequences between positions -222 and -167 were required for efficient expression in primary embryonic chick lens cells; sequences downstream of the poly(A) addition signal were dispensable for expression in this in vitro system.