Batzer M A, Desselle T D, Brennan M D, Lee W R, Tedeschi B
Department of Zoology and Physiology, Louisiana State University, Baton Rouge 70803.
Biochem Biophys Res Commun. 1988 Jan 29;150(2):655-64. doi: 10.1016/0006-291x(88)90442-1.
The protein expressed by the alcohol dehydrogenase locus (Adh) in D. melanogaster comprises a small group of electromorphs. We are able to study the expression of these electromorphs by electrophoretic separation and subsequent probing of blots of the separated polypeptides with antiserum for alcohol dehydrogenase (ADH). In the present study we have utilized this technique to study and compare the ADH electromorphs in wild type D. melanogaster with D. melanogaster transformants which carry an Adh gene from D. grimshawi, D. hawaiiensis or D. affinidisjuncta and produced functional ADH (10, 19). We have determined that polypeptides are produced by the donor loci in the transformed flies and further show that although the molecular weight of the expressed polypeptides is similar to D. melanogaster electromorphs, the isoelectric points are not similar. Thus this methodology offers the potential to study naturally occurring ADH electromorphs and null alleles independent of enzymatic activity assays.
黑腹果蝇中酒精脱氢酶基因座(Adh)所表达的蛋白质包含一小群电泳变体。我们能够通过电泳分离,随后用抗酒精脱氢酶(ADH)血清对分离出的多肽印迹进行探测,来研究这些电泳变体的表达情况。在本研究中,我们利用该技术对野生型黑腹果蝇与携带来自格氏果蝇、夏威夷果蝇或亲缘间断果蝇的Adh基因并产生功能性ADH的黑腹果蝇转化体中的ADH电泳变体进行研究和比较(10, 19)。我们已经确定转化果蝇中的供体基因座会产生多肽,并且进一步表明,尽管所表达多肽的分子量与黑腹果蝇的电泳变体相似,但其等电点并不相似。因此,这种方法提供了独立于酶活性测定来研究天然存在的ADH电泳变体和无效等位基因的潜力。