Bansal K C, Viret J F, Haley J, Khan B M, Schantz R, Bogorad L
Department of Cellular and Developmental Biology, Harvard University, Cambridge, MA 02138.
Proc Natl Acad Sci U S A. 1992 Apr 15;89(8):3654-8. doi: 10.1073/pnas.89.8.3654.
Cell-specific and light-regulated expression of the beta-glucuronidase (GUS) reporter gene from maize cab-m1 and rbcS-m3 promoter sequences was studied in maize leaf segments by using an in situ transient expression microprojectile bombardment assay. The cab-m1 gene is known to be strongly photoregulated and to be expressed almost exclusively in mesophyll cells (MC) but not in bundle sheath cells (BSC). Expression of GUS from a 1026-base-pair 5' promoter fragment of cab-m1 is very low in dark-grown leaves; GUS expression is increased about 10-fold upon illumination of dark-grown leaves. In illuminated leaves, the ratio of GUS expression in MC vs. BSC is about 10:1. The cab-m1 region between 868 and 1026 base pairs 5' to the translation start confers strong MC-preferred expression on the remainder of the chimeric gene in illuminated leaves, but a region between -39 and -359 from the translation start is required for photoregulated expression. Transcripts of rbcS-m3 are found in BSC but not in MC and are about double in BSC of greening dark-grown seedlings. In contrast to the behavior of the cab-m1-GUS construct, GUS expression driven by 2.1 kilobase pairs of the rbcS-m3 5' region was about twice as high in MC as in BSC of unilluminated dark-grown maize leaves. The number of BSC, but not MC, expressing GUS nearly doubled upon greening of bombarded etiolated leaves. These data suggest that the 5' region of rbcS-m3 used here could be responsible for most of the light-dependent increase in rbcS-m3 transcripts observed in BSC of greening leaves and that transcriptional or posttranscriptional mechanisms are responsible for the lack of rbcS-m3 transcripts in MC.
利用原位瞬时表达微弹轰击试验,研究了来自玉米cab - m1和rbcS - m3启动子序列的β - 葡萄糖醛酸酶(GUS)报告基因在玉米叶片切段中的细胞特异性和光调节表达。已知cab - m1基因受强光调节,几乎只在叶肉细胞(MC)中表达,而不在维管束鞘细胞(BSC)中表达。来自cab - m1的1026个碱基对的5'启动子片段驱动的GUS在黑暗生长的叶片中表达非常低;黑暗生长的叶片光照后,GUS表达增加约10倍。在光照叶片中,MC与BSC中GUS表达的比例约为10:1。翻译起始位点5'端868至1026个碱基对之间的cab - m1区域赋予嵌合基因其余部分在光照叶片中强烈的MC优先表达,但翻译起始位点 - 39至 - 359之间的区域是光调节表达所必需的。rbcS - m3的转录本在BSC中发现,而不在MC中,并且在黑暗生长的绿化幼苗的BSC中约为两倍。与cab - m1 - GUS构建体的行为相反,由rbcS - m3 5'区域的2.1千碱基对驱动的GUS表达在未光照的黑暗生长玉米叶片的MC中比在BSC中高约两倍。轰击的黄化叶片绿化后,表达GUS的BSC数量几乎增加了一倍,但MC没有增加。这些数据表明,这里使用的rbcS - m3的5'区域可能是绿化叶片BSC中观察到的rbcS - m3转录本光依赖性增加的主要原因,并且转录或转录后机制是MC中缺乏rbcS - m3转录本的原因。