旋毛虫与伪旋毛虫肌幼虫抗原的SDS-PAGE和双向电泳分析

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。

Comparative analysis of muscle larval antigens of Trichinella spiralis and Trichinella pseudospiralis by SDS-PAGE and two-dimensional gel electrophoresis

To identify the different proteins of muscle larvae of Trichinella spiralis （T1） and T. pseudospiralis （T4）, The soluble antigen and excretory-secretory （ES） antigens of T1 and T4 muscle larvae cultured in vitro for 24 h were analyzed by SDS-PAGE and two dimensional gel electrophoresis （2-DE）. The results of SDS-PAGE demonstrate that T1 muscle larval soluble antigens have 22 protein bands（221.62--14.88 kDa） , 6 bands of which（59. 72 kDa,44. 37 kDa,23. 66 kDa,22.36 kDa,18.26 kDa,16.34 kDa）are T1 specific; T4 shows 18 protein bands （185. 28-- 14. 27 kDa） with 4 bands （132. 60 kDa, 119.30 kDa, 35. 26 kDa,31. 02 kDa） being T4-specific. The T1 muscle larval ES antigens show 10 bands（ll3. 21--14. 37 kDa）, while T4 have 9 bands（104.71--14.51 kDa）. All the protein bands of T1 and T4 muscle larval ES antigens are differ ent. The results of 2-DE reveal that T1 muscle larval soluble antigens have 193--4-12 protein spots mainly located in the molecular weight areas of 11-- 22 kDa, 25-- 64 kDa and 100 -- 144 kDa, the corresponding isoelectric point （pH） is 4.7 -- 8.2,4.5 -- 6.5and 5--7; T4 have 175±9 protein points mostly located in the areas of 12--21 kDa and 25--90 kDa, the corresponding pI is 4--9.5 and 4.5--9.6. T1 muscle larval ES antigens have 82±6 protein spots mainly located in the areas of 13--16 kDa,18 22 kDa and 40--55 kDa, the pI is 4--7,3.8--6.2 and 5--9; The T4 ES antigens represent 69±5 spots mainly in the areas of 10--15 kDa,17--25 kDaand 29--55 kDa, the pI is 4.7 6. 5, 4. 6--6 and 5 7. In conclusion, the protein components in muscle larval soluble and ES antigens of T. spiralis are obviously different from that of T. pseudospiralis.