McKeown M, Firtel R A
Cell. 1981 Jun;24(3):799-807. doi: 10.1016/0092-8674(81)90105-7.
Using a modification of the Berk and Sharp S1 nuclease mapping procedure and by analyzing actin cDNA clones, we have examined the expression of several members of the 17-member multigene family encoding actin in Dictyostelium. The mapping procedure, which takes advantage of the fact that the actin genes are homologous in the protein-coding region but are very divergent in the proposed 5' untranslated region has enabled us to quantitate the relative expression of several genes during the Dictyostelium life cycle. We have shown that at least six of the 17 potential actin-coding sequences are expressed. One is not expressed at levels of more than 0.5--1% of total actin mRNA at the developmental times examined and appears to be a pseudogene. By quantitating the amount of actin mRNA in mRNA populations isolated from cells at various times in development, we have shown that four of the actin genes show different patterns of expression. Interestingly, three of the four genes appear to encode the same protein. We have also taken advantage of the S1 mapping procedure to identify the 5' ends of the actin mRNAs from four genes and have compared the sequences outside the 5' ends on these genes with the nucleotide sequences of seven other actin genes. We have identified homologous sequences in most of these genes that may be involved in initiation of transcription.
通过对伯克和夏普S1核酸酶图谱分析方法进行改进,并对肌动蛋白cDNA克隆进行分析,我们研究了盘基网柄菌中17个成员的多基因家族中几个编码肌动蛋白成员的表达情况。该图谱分析方法利用了肌动蛋白基因在蛋白质编码区域具有同源性,但在假定的5'非翻译区域差异很大这一事实,使我们能够定量盘基网柄菌生命周期中几个基因的相对表达。我们已经表明,17个潜在的肌动蛋白编码序列中至少有6个表达。在所检测的发育阶段,其中一个基因的表达水平不超过总肌动蛋白mRNA的0.5%-1%,似乎是一个假基因。通过定量从发育不同阶段的细胞中分离的mRNA群体中肌动蛋白mRNA的量,我们发现其中四个肌动蛋白基因表现出不同的表达模式。有趣的是,这四个基因中的三个似乎编码相同的蛋白质。我们还利用S1图谱分析方法鉴定了四个基因的肌动蛋白mRNA的5'末端,并将这些基因5'末端以外的序列与其他七个肌动蛋白基因的核苷酸序列进行了比较。我们在大多数这些基因中鉴定出了可能参与转录起始的同源序列。