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[通过丙烯酰胺凝胶电泳对猪支原体进行鉴别(作者译)]

[The differenziation of swine-mycoplasma by acrylamide-gel-electrophoresis (author's transl)].

作者信息

Balke E

出版信息

Zentralbl Bakteriol Orig A. 1975;230(3):398-405.

PMID:49991
Abstract

The strains of swine mycoplasma (Tab. 1) were grown aerobically in Whittlestone's Medium containing swine serum for 4-5 days at 37 degrees C. After centrifugation and washing they were freeze-dried and extracted with a phenol-acetic acid-water solution (4:2:1) (1 ml/5 mg mycoplasma dru weight) at 4 degrees C over 48 h. After centrifugation 2 volumes of the supernatant were mixed with 1 volume of a 40% sucrose solution in 35% acetic acid. 0,15 or 0,2 ml of which were added to the acrylamide-gel (2 ml in a tube 0,5 times 10 cm) containing 5 M urea and 35% acetic acid. This solution was overlayed with 0,1 ml 75% acetic acid and tubes then filled with 10% acetic acid. The solution in both electrode chambers was 10% acetic acid, too. During the first 5 min it was separated with 2 mA/tube, then 3 or 5 h with 4 mA/tube. The protein bands were stained with amido black 10B. For control steril culture medium was investigated, too. Various preparations of freeze-dried M. hyopneumoniae gave identical protein patterns. Nearly identical were the patterns of protein bands from M. suipneumoniae and M. hyopneumoniae; that means identity of the strains or very close relationship (Figs. 1 and 2). This is in agreement with other authors who investigated both strains with serological methods. M. suipneumoniae and M. hyopneumoniae were found to be different as well from M. hyorhinis and M. sp. E9 from M. granularum (Figs. 1 and 2). Between M. hyorhinis and M. sp. E9 less relationship was noted. All these results were in agreement with investigations performed with the aid of Latex agglutination.

摘要

猪支原体菌株(表1)在含有猪血清的惠特尔斯通培养基中于37℃需氧培养4 - 5天。离心洗涤后,将它们冻干,并在4℃下用酚 - 乙酸 - 水溶液(4:2:1)(1 ml/5 mg支原体干重)提取48小时。离心后,将2倍体积的上清液与1倍体积的40%蔗糖溶液在35%乙酸中混合。取0.15或0.2 ml加入到含有5 M尿素和35%乙酸的丙烯酰胺凝胶(0.5×10 cm管中2 ml)中。该溶液用0.1 ml 75%乙酸覆盖,然后管中充满10%乙酸。两个电极室中的溶液也为10%乙酸。最初5分钟以2 mA/管进行分离,然后3或5小时以4 mA/管进行分离。蛋白质条带用氨基黑10B染色。同时也对对照无菌培养基进行了研究。冻干的猪肺炎支原体的各种制剂给出了相同的蛋白质图谱。猪肺炎支原体和猪肺炎支原体的蛋白质条带图谱几乎相同;这意味着菌株相同或关系非常密切(图1和图2)。这与其他用血清学方法研究这两种菌株的作者的结果一致。发现猪肺炎支原体和猪肺炎支原体也与猪鼻支原体和颗粒支原体的E9菌株不同(图1和图2)。猪鼻支原体和E9菌株之间的关系较弱。所有这些结果与借助乳胶凝集试验进行的研究结果一致。

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