Aly R, Granot D, Mahler-Slasky Y, Halpern N, Nir D, Galun E
Department of Weed Research, A.R.O., Newe Ya'ar Research Center, Ramat Yishay, 30095, Israel.
Protein Expr Purif. 1999 Jun;16(1):120-4. doi: 10.1006/prep.1999.1059.
Sarcotoxin IA is a cecropin-type antibacterial protein produced by the flesh fly, Sarcophaga peregrina. Similar to other bactericidal small proteins produced by insects, sarcotoxin IA is released into the hemolymph of larvae and nymphs upon mechanical injury or bacterial infection. The gene (sarco) that encodes this toxin was introduced into Saccharomyces cerevisiae yeast cells and was expressed under a constitutive yeast promoter. The transformed yeast cells were grown in a liquid medium, and a peptide with a similar molecular size to that of the mature sarcotoxin IA was detected in the medium by Western blot analysis. The secreted sarcotoxin-like peptide (SLP) had a potent cytotoxic effect against several bacteria, including plant pathogenic bacteria, similar to the toxic effects of the authentic sarcotoxin IA. Erwinia carotovora was more susceptible to the toxic medium than Pseudomonas solanacearum and Pseudomonas syringae pv. lachrymans. Thus, yeast may be used in the production of such proteins for employment against various bacterial pathogens.
肌毒素IA是一种由麻蝇(Sarcophaga peregrina)产生的天蚕素型抗菌蛋白。与昆虫产生的其他杀菌小蛋白类似,肌毒素IA在受到机械损伤或细菌感染时会释放到幼虫和若虫的血淋巴中。编码这种毒素的基因(sarco)被导入酿酒酵母细胞中,并在组成型酵母启动子的控制下表达。将转化后的酵母细胞在液体培养基中培养,通过蛋白质免疫印迹分析在培养基中检测到一种与成熟肌毒素IA分子大小相似的肽。分泌的肌毒素样肽(SLP)对包括植物病原菌在内的几种细菌具有强大的细胞毒性作用,类似于天然肌毒素IA的毒性作用。胡萝卜软腐欧文氏菌比青枯假单胞菌和丁香假单胞菌番茄致病变种更易受到有毒培养基的影响。因此,酵母可用于生产此类蛋白质以对抗各种细菌病原体。