Méténier L, Kaminski M
Ann Immunol (Paris). 1978 Feb-Mar;129(2-3):353-65.
Immunochemical and enzymatic analyses of horse serum carboxylesterase were carried out with respect to the existence of a silent gene. Sera with positive phenotypic expression of esterase, both heterozygotes and presumed homozygotes, were compared with:--sera with positive phenotypic expression but genotypically +/O;--sera with a negative phenotypic expression, i. e. genotypically O/O;--sera of natural +/O "hemi-zygotes": mules (donkey lacking the esterase);--positive sera heated at 60 degrees C;--positive sera after specific inhibition of enzymatic activity. Titration by immunocompetition has shown that sera phenotypically negative do not contain an immunologically active "dummy" protein; however, using a sensitive immunotitration (rocket immunoelectrophoresis), traces of active protein could be demonstrated. Sera of +/O genotypes, both horses and mules, contain approximatively half of esterase present in +/+ sera. Heating at 60 degrees destroys the antigenic properties of the esterase almost completely, while, on the contrary, a specific inhibition of enzymatic activity leaves the antigenic properties unaffected.
针对沉默基因的存在,对马血清羧酸酯酶进行了免疫化学和酶学分析。将具有酯酶阳性表型表达的血清(包括杂合子和假定的纯合子)与以下血清进行比较:——具有阳性表型表达但基因型为+/O的血清;——具有阴性表型表达(即基因型为O/O)的血清;——天然+/O“半合子”血清:骡子(缺乏酯酶的驴);——在60℃加热的阳性血清;——酶活性被特异性抑制后的阳性血清。免疫竞争滴定表明,表型阴性的血清不含有免疫活性的“假”蛋白;然而,使用灵敏的免疫滴定法(火箭免疫电泳),可以检测到微量的活性蛋白。+/O基因型的血清,包括马和骡子的血清,所含酯酶约为+/+血清中酯酶的一半。60℃加热几乎完全破坏了酯酶的抗原特性,相反,酶活性的特异性抑制对抗原特性没有影响。