Rogers M V, Henkle K J, Herrmann V, McLaren D J, Mitchell G F
Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria, Australia.
Infect Immun. 1991 Apr;59(4):1442-7. doi: 10.1128/iai.59.4.1442-1447.1991.
Type A2 phospholipase (PLA2) activity has been observed in integral membrane protein extracts of Schistosoma japonicum. Antiserum raised against bee venom PLA2 recognized a single 16-kDa band in the parasite extracts; it also localized to antigen in the gut lining of fixed adult schistosomes as shown by immunofluorescence techniques. Evidence was obtained that the molecule was expressed at low levels in comparison with other integral membrane proteins and was weakly immunogenic in rabbits. Two oligonucleotide probes were constructed on the basis of highly conserved regions between the nucleotide sequences of rat, bovine, rattlesnake, and bee venom PLA2; these probes were used to isolate S. japonicum genomic DNA phage clones. A 1.8-kb FnuD2 fragment was shown by Southern blot analysis to strongly hybridize with the 5' 32P-labeled PLA2 oligonucleotides in both S. japonicum genomic DNA and DNA from one of the phage clones. The nucleotide and predicted amino acid sequences of this fragment revealed homology with the C terminus of PLA2s from different species.
在日本血吸虫的整合膜蛋白提取物中观察到了A2型磷脂酶（PLA2）活性。用抗蜂毒PLA2产生的抗血清在寄生虫提取物中识别出一条单一的16 kDa条带；通过免疫荧光技术显示，它也定位于固定的成年血吸虫肠壁中的抗原。有证据表明，与其他整合膜蛋白相比，该分子表达水平较低，并且在兔中免疫原性较弱。基于大鼠、牛、响尾蛇和蜂毒PLA2核苷酸序列之间的高度保守区域构建了两个寡核苷酸探针；这些探针用于分离日本血吸虫基因组DNA噬菌体克隆。通过Southern印迹分析表明，一个1.8 kb的FnuD2片段在日本血吸虫基因组DNA和来自其中一个噬菌体克隆的DNA中均与5' 32P标记的PLA2寡核苷酸强烈杂交。该片段的核苷酸和预测的氨基酸序列显示与来自不同物种的PLA2的C末端具有同源性。
