Tumer N E, Clark W G, Tabor G J, Hironaka C M, Fraley R T, Shah D M
Nucleic Acids Res. 1986 Apr 25;14(8):3325-42. doi: 10.1093/nar/14.8.3325.
We have isolated five members of the multigene family encoding the small subunit (rbcS) of ribulose-1,5-bisphosphate carboxylase in petunia and examined their expression in petunia leaves. Of the five rbcS genes, two (ssu11A and ssu8) are expressed at high levels in petunia leaves. Northern analysis using gene specific oligonucleotide probes revealed that ssu11A accounts for 40% of the total rbcS transcripts in petunia leaves while ssu8 accounts for 4 to 5% of the total rbcS transcripts. Structural comparisons of ssu8 and ssu11A revealed that the coding sequence of ssu8 is interrupted by three introns, while the coding sequence of ssu11A is interrupted by two introns. The positions of the first two introns are identical, the third intron in ssu8 is located in a highly conserved region of the protein. The 5' and 3' flanking sequences of ssu11A are highly homologous to the 5' and 3' flanking sequences of ssu8. S1 nuclease mapping was used to locate the start of transcription of ssu8 and ssu11A and showed that ssu8 mRNA leader differs in sequence from the ssu11A mRNA leader.
我们从矮牵牛中分离出了编码1,5-二磷酸核酮糖羧化酶小亚基(rbcS)的多基因家族的五个成员,并检测了它们在矮牵牛叶片中的表达情况。在这五个rbcS基因中,有两个(ssu11A和ssu8)在矮牵牛叶片中高水平表达。使用基因特异性寡核苷酸探针进行的Northern分析表明,ssu11A占矮牵牛叶片中rbcS转录本总量的40%,而ssu8占rbcS转录本总量的4%至5%。ssu8和ssu11A的结构比较显示,ssu8的编码序列被三个内含子打断,而ssu11A的编码序列被两个内含子打断。前两个内含子的位置相同,ssu8中的第三个内含子位于蛋白质的高度保守区域。ssu11A的5'和3'侧翼序列与ssu8的5'和3'侧翼序列高度同源。使用S1核酸酶作图来定位ssu8和ssu11A的转录起始点,结果表明ssu8 mRNA的前导序列在序列上与ssu11A mRNA的前导序列不同。