Gurevitz M, Zlotkin E, Zilberberg N
Department of Botany, Tel-Aviv University, Israel.
FEBS Lett. 1990 Aug 20;269(1):229-32. doi: 10.1016/0014-5793(90)81161-g.
The poly(A)+ mRNA isolated from the venomous terminal segments of the scorpion Buthotus judaicus was reversed transcribed into cDNA. PCR amplification of the cDNA in presence of oligonucleotide primers prepared on basis of the known amino acid sequence of the depressant insect toxin II yielded a 125 bp long product. This fragment was cloned and its sequence determined. The deduced amino acid sequence has revealed a complete homology with the amino acid sequence of the toxin. This clone was used to probe a Northern blot resolving the poly(A)+ and poly(A)- fractions derived from the scorpion. An organ specific 360 nucleotide transcript which might be the processed product of a approximately 4.0 kb precursor was elucidated. This cDNA clone may pave the way for a molecular genetic approach to study the structure-function relationship of scorpion selective insect toxins.
从犹大杀牛蝎(Buthotus judaicus)有毒的尾部节段分离出的聚腺苷酸加尾(poly(A)+)mRNA被逆转录成cDNA。在根据抑制性昆虫毒素II已知氨基酸序列制备的寡核苷酸引物存在的情况下,对该cDNA进行PCR扩增,得到了一个125 bp长的产物。该片段被克隆并测定了其序列。推导的氨基酸序列显示与该毒素的氨基酸序列完全同源。这个克隆被用来探测Northern印迹,解析来自蝎子的聚腺苷酸加尾(poly(A)+)和非聚腺苷酸加尾(poly(A)-)组分。阐明了一种器官特异性的360个核苷酸的转录本,它可能是一个约4.0 kb前体的加工产物。这个cDNA克隆可能为研究蝎子选择性昆虫毒素的结构-功能关系的分子遗传学方法铺平道路。