目的鉴定旋毛虫(Trichinella spiralis,T1)与伪旋毛虫(T.pseudospiralis,T4)肌幼虫的差异蛋白。方法应用SDS-PAGE和双向电泳(two-dimensional gel electrophoresis,2-DE)对T1、T4肌幼虫的可溶性抗原与培养24h的ES抗原的蛋白组分进行分...目的鉴定旋毛虫(Trichinella spiralis,T1)与伪旋毛虫(T.pseudospiralis,T4)肌幼虫的差异蛋白。方法应用SDS-PAGE和双向电泳(two-dimensional gel electrophoresis,2-DE)对T1、T4肌幼虫的可溶性抗原与培养24h的ES抗原的蛋白组分进行分析。结果SDS-PAGE显示,T1肌幼虫可溶性抗原有22条蛋白带(221.62kDa～14.88kDa),其中6条为T1特异性蛋白带(59.72、44.37、23.66、22.36、18.26、16.34kDa);T4可溶性抗原蛋白有18条带(185.28kDa～14.27kDa),其中4条为T4特异性蛋白带(132.60、119.30、35.26、31.02kDa)。T1的ES抗原有10条蛋白带(113.21kDa～14.37kDa),T4的ES抗原有9条蛋白带(104.71kDa～14.51kDa),T1、T4肌幼虫ES抗原的蛋白带均不相同。2-DE显示,T1可溶性抗原有193±12个蛋白点,分子量主要为11kDa～22kDa、25kDa～64kDa及100kDa～144kDa,所对应的等电点(pI)分别为4.7～8.2、4.5～6.5及5～7;T4可溶性抗原有175±9个蛋白点,分子量主要为12kDa～21kDa及25kDa～90kDa,所对应的pI分别为4～9.5与4.5～9.6。T1的ES抗原具有82±6个蛋白点,分子量主要为13kDa～16kDa、18kDa～22kDa及40kDa～55kDa,所对应的pI分别为4～7、3.8～6.2及5～9;T4的ES抗原具有69±5个蛋白点,分子量主要为10kDa～15kDa、17kDa～25kDa及29kDa～55kDa,所对应的pI分别为4.7～6.5、4.6～6及5～7。结论旋毛虫肌幼虫可溶性抗原及ES抗原的蛋白组分与伪旋毛虫的明显不同。展开更多
An improved CBB staining with higher sensitivity than that of the typical CBB staining was reported.The main improvement was using a fixing step of 25% trichloroacetic acid(TCA) before CBB staining.For most proteins s...An improved CBB staining with higher sensitivity than that of the typical CBB staining was reported.The main improvement was using a fixing step of 25% trichloroacetic acid(TCA) before CBB staining.For most proteins studied,the sensitivity of the improved CBB staining was about twice as high as that of the typical method.For basic and low molecular weight proteins such as ribosomal proteins,the sensitivity of this improved staining method was about 3.5-28 times that of the typical method.It was speculated that the improved procedure would be suitable for exact quantitative analysis of proteins fractionated by SDS-PAGE,especially for basic and low molecular weight proteins.On the other hand,this new modified method might be also applied to multidisciplinary studies,such as biological researches and nuclear sciences.展开更多
文摘目的鉴定旋毛虫(Trichinella spiralis,T1)与伪旋毛虫(T.pseudospiralis,T4)肌幼虫的差异蛋白。方法应用SDS-PAGE和双向电泳(two-dimensional gel electrophoresis,2-DE)对T1、T4肌幼虫的可溶性抗原与培养24h的ES抗原的蛋白组分进行分析。结果SDS-PAGE显示,T1肌幼虫可溶性抗原有22条蛋白带(221.62kDa～14.88kDa),其中6条为T1特异性蛋白带(59.72、44.37、23.66、22.36、18.26、16.34kDa);T4可溶性抗原蛋白有18条带(185.28kDa～14.27kDa),其中4条为T4特异性蛋白带(132.60、119.30、35.26、31.02kDa)。T1的ES抗原有10条蛋白带(113.21kDa～14.37kDa),T4的ES抗原有9条蛋白带(104.71kDa～14.51kDa),T1、T4肌幼虫ES抗原的蛋白带均不相同。2-DE显示,T1可溶性抗原有193±12个蛋白点,分子量主要为11kDa～22kDa、25kDa～64kDa及100kDa～144kDa,所对应的等电点(pI)分别为4.7～8.2、4.5～6.5及5～7;T4可溶性抗原有175±9个蛋白点,分子量主要为12kDa～21kDa及25kDa～90kDa,所对应的pI分别为4～9.5与4.5～9.6。T1的ES抗原具有82±6个蛋白点,分子量主要为13kDa～16kDa、18kDa～22kDa及40kDa～55kDa,所对应的pI分别为4～7、3.8～6.2及5～9;T4的ES抗原具有69±5个蛋白点,分子量主要为10kDa～15kDa、17kDa～25kDa及29kDa～55kDa,所对应的pI分别为4.7～6.5、4.6～6及5～7。结论旋毛虫肌幼虫可溶性抗原及ES抗原的蛋白组分与伪旋毛虫的明显不同。
文摘An improved CBB staining with higher sensitivity than that of the typical CBB staining was reported.The main improvement was using a fixing step of 25% trichloroacetic acid(TCA) before CBB staining.For most proteins studied,the sensitivity of the improved CBB staining was about twice as high as that of the typical method.For basic and low molecular weight proteins such as ribosomal proteins,the sensitivity of this improved staining method was about 3.5-28 times that of the typical method.It was speculated that the improved procedure would be suitable for exact quantitative analysis of proteins fractionated by SDS-PAGE,especially for basic and low molecular weight proteins.On the other hand,this new modified method might be also applied to multidisciplinary studies,such as biological researches and nuclear sciences.
