Alcaino H A, Baker N F, Fisk R A
Am J Vet Res. 1976 Oct;37(10):1153-7.
Homogenates of individual adult Fasciola hepatica, collected from bile ducts of sheep, were subjected to isoelectric focusing in polyacrylamide flat-bed gels after which zymograms were made by enzymatic and staining reactions. Seven identifiable isozymes that hydrolyzed alpha-naphthyl acetate were found. These 7 isozymes were identified by their isoelectric points as 5.10, 5.15, 5.25, 5.40, 5.55, 5.65, and 5.75. None of these isozymes was common to all F hepatica examined. Combinations of these isozymes in individual flukes revealed 7 distinct phenotypes, the frequency of which varied between local populations, i.e., populations of flukes obtained from individual sheep of different origin. In addition to these 7 isozymes, there were weak staining bands of enzyme activity, between pH 5.8 and the cathode end of the gels, which could not be accurately identified. The method of isoelectric focusing and zymogram preparation used in the present study did not reveal cholinesterase activity, even though the presence of activity in homogenates was demonstrated by polyacrylamide disc-gel electrophoresis. Strong activity of acid phosphatase between pH 5 and 6 was found, but poor resolution prevented obtaining accurate identification of isozymes. Weak activity with poor resolution of isozymes of alkaline phosphatase was observed between pH 4.4 and 5.75.
从绵羊胆管采集的成年肝片吸虫个体匀浆，在聚丙烯酰胺平板凝胶中进行等电聚焦，然后通过酶促反应和染色反应制作酶谱图。发现了七种可识别的水解α-萘乙酸的同工酶。根据等电点，这七种同工酶分别为5.10、5.15、5.25、5.40、5.55、5.65和5.75。在所检测的所有肝片吸虫中，没有一种同工酶是共同存在的。个体吸虫中这些同工酶的组合显示出7种不同的表型，其频率在不同的本地种群（即从不同来源的个体绵羊获得的吸虫种群）之间有所变化。除了这七种同工酶外，在pH 5.8至凝胶阴极末端之间存在酶活性的弱染色带，无法准确识别。本研究中使用的等电聚焦和酶谱图制备方法未显示出胆碱酯酶活性，尽管聚丙烯酰胺圆盘凝胶电泳证明匀浆中存在该活性。在pH 5至6之间发现了较强的酸性磷酸酶活性，但分辨率较差，无法准确鉴定同工酶。在pH 4.4至5.75之间观察到碱性磷酸酶同工酶活性较弱且分辨率较差。
