Use of P-element-mediated transformation to identify the molecular basis of naturally occurring variants affecting Adh expression in Drosophila melanogaster.
Use of P-element-mediated transformation to identify the molecular basis of naturally occurring variants affecting Adh expression in Drosophila melanogaster.
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使用 P 元件介导的转化来鉴定影响果蝇 Adh 表达的自然发生变异的分子基础。
DOI：
10.1093/genetics/115.1.129
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发表时间：
1987
期刊：
影响因子：
3.3
通讯作者：
Stam,LF
中科院分区：
文献类型：
--
作者：
Laurie-Ahlberg,CC;Stam,LF
本研究旨在探讨果蝇乙醇脱氢酶(ADH)基因表达水平差异的分子基础。以前的研究表明，快速品系的活性水平通常是慢速品系的两到三倍，而且它们的ADH蛋白水平(从免疫学上估计)也要高得多。一项与ADH活性变化有关的限制性片段长度多态性研究结果表明，等位酶之间Adh表达的差异可能不完全是由于氨基酸替代，而可能部分是由于与调控位点多态性的连锁不平衡。在这里，我们描述了一种利用OFP元件介导的转化来识别导致ADH水平差异的核苷酸替换(S)的方法。这种方法包括在来自典型的慢/快等位基因对的体外培养的两个粘附区克隆中产生重组，然后测试特定的限制性片段对活体转化表达的影响。利用这种方法，对ADH活性和ADH蛋白水平的影响明显地映射到一个2.3kb的限制性片段，该片段包括所有的Adhcoding序列和一些内含子和3‘侧翼序列，但排除了远端(成体)转录单位的所有5’侧翼序列。Kreitman对这一片段的DNA序列与几个Slow和Fast等位基因进行了比较，显示出活性水平的典型差异，表明只有三个核苷酸替换将所有Fast与所有Slow等位基因区分开来。因此，很可能是这些替换中的一个或多个导致了不同等位酶类之间Adh表达的主要差异。这些替换中的一个当然是慢/快氨基酸替换替换(在1490)，而另外两个是附近的第三位沉默替换(在1443和1527)。对转化子群体间差异的定量分析表明，元素转化法可用于定位对基因表达的相对较小的影响(约20%)。
The purpose of the work reported here is to identify the molecular basis of the difference in level of expression between the polymorphic Slow and Fast alcohol dehydrogenase (Adh) alleles inDrosophila melanogaster. Previous studies have shown that Fast lines typically have a two- to threefold higher activity level than Slow lines and they also have a substantially higher level of ADH-protein (estimated immunologically). The results of a restriction fragment length polymorphism study in relation to ADH activity variation had previously suggested that the difference inAdhexpression between allozymes might not be due entirely to the amino acid replacement substitution, but could be due in part to linkage disequilibrium with a regulatory site polymorphism. Here we describe an approach that makes use ofP-element-mediated transformation in order to identify the nucleotide substitution(s) responsible for this difference in ADH level. This approach consists of generating recombinantsin vitrobetweenAdhregion clones derived from a typical Slow/Fast pair of alleles and then testing for the effects of particular restriction fragments on expressionin vivoby transformation. Using this approach, the effect on both ADH activity and ADH-protein level clearly maps to a 2.3-kb restriction fragment that includes all of theAdhcoding sequence and some intron and 3' flanking sequence, but excludes all of the 5' flanking sequence of the distal (adult) transcriptional unit. Comparison of Kreitman's DNA sequences for this fragment from several Slow and Fast alleles showing the typical difference in activity level shows that only three nucleotide substitutions distinguish all Fast from all Slow alleles. Thus, it is likely that one or more of these substitutions causes the major difference inAdhexpression between allozymic classes. One of these substitutions is, of course, the Slow/Fast amino acid replacement substitution (at 1490) while the other two are nearby third position silent substitutions (at 1443 and 1527). A quantiative analysis of variation among transformant stocks shows that theP-element transformation approach can be used to localize even relatively small effects on gene expression (on the order of 20%).
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果蝇中 Adh 表达的遗传控制。
影响因子：
3.3
作者：
G. Maroni;C. C. Laurie
通讯作者：
C. C. Laurie
果蝇胶蛋白基因 Sgs4 附近相互作用的 DNAase I 超敏感位点的复合体
影响因子：
64.5
作者：
A. W. Shermoen;S. Beckendorf
通讯作者：
S. Beckendorf
DOI：
--
发表时间：
1980
期刊：
Basic life sciences
影响因子：
--
作者：
Arthur Chovnick;M. McCarron;Stephen H. Clark;A. Hilliker;Christine Rushlow
通讯作者：
Christine Rushlow
影响因子：
3.8
作者：
A. Birley;P. Couch;A. Marson
通讯作者：
A. Marson
果蝇玫瑰色基因座的组织：支持与黄嘌呤脱氢酶结构元件相邻的顺式作用控制元件的进一步证据。
影响因子：
3.3
作者：
M. McCarron;J. O'donnell;A. Chovnick;B. S. Bhullar;J. Hewitt;E. Candido
通讯作者：
E. Candido