Rössler H, Burkhardt H, Rosenthal S, Scholz D, Rosenthal H A, Altaner C
Folia Biol (Praha). 1985;31(4):273-83.
The permanently BLV-infected FLK cell line 44/2 and FLK sublines were tested for the stability of their BLV and antigen synthesis by three virus markers, p24, gp51, and activity of reverse transcriptase. The extent of BLV production in the FLK line correlated directly with the surface for cell growth in roller and stationary cultures. The synthesis and secretion of non-virus-associated gp51 is especially stimulated in the roller culture, and is largely independent of the quality of the culture medium. The roller culture allows considerable economy of the medium, at the same time retaining its full biological activity. As much as 2 mg of gp51 per litre of culture supernatant was obtained by this procedure. Appropriate markers for estimation of BLV production are p24 and gp51. For this purpose, the activity of reverse transcriptase on its own was not sufficient. The BLV yield differed by one order of magnitude among the 18 cloned FLK sublines. Three sublines have been identified, which showed a comparatively high virus production, under conditions of stationary cultures. The amount or activity of viral markers could not be correlated with the number of BLV proviruses.
利用三种病毒标志物p24、gp51和逆转录酶活性,对永久感染牛白血病病毒(BLV)的FLK细胞系44/2及其亚系的BLV稳定性和抗原合成进行了检测。FLK细胞系中BLV的产生程度与滚瓶培养和平板培养中的细胞生长面积直接相关。非病毒相关gp51的合成和分泌在滚瓶培养中尤其受到刺激,并且在很大程度上与培养基的质量无关。滚瓶培养可显著节省培养基,同时保持其全部生物活性。通过该方法,每升培养上清液可获得多达2mg的gp51。用于评估BLV产生的合适标志物是p24和gp51。为此,仅逆转录酶活性是不够的。在18个克隆的FLK亚系中,BLV产量相差一个数量级。已鉴定出三个亚系,在平板培养条件下表现出相对较高的病毒产生量。病毒标志物的数量或活性与BLV前病毒的数量无关。