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噬菌体T4中的DNA复制与头部组装

DNA replication and head assembly in bacteriophage T4.

作者信息

Andreev V M, Gonikberg E M

出版信息

Mol Biol (Mosk). 1976 Jul-Aug;10(4):685-96.

PMID:1023049
Abstract

Phage DNA was accumulated in cells of E. coli B, infected with the phage T4DtsLB3 (gene 42), without the synthesis of late proteins (in the presence of chloramphenicol). Then (stage II), chloramphenicol was removed and further replication of the phage DNA suppressed with hydroxyurea and by simultaneously raising the temperature to 40 degrees. The media M9 or M9 with 1% amino acid were used; the times of addition of chloramphenicol and the hydroxyurea concentration were also varied. It was also shown that in medium M9, at stage II, chiefly early proteins were synthesized. In the medium containing amino acids, at stage II the following was observed: 1) DNA synthesis was entirely suppressed and a degradation of DNA occurred; 2) both early and late proteins were synthesized, with a predominance of the latter; 3) an assembly of the elements of the phage tails and capsids occurred without the neck and flagellum, and a small number of phage particles were also found; 4) the capsids, isolated in a sucrose density gradient after lysis with chloroform, contained the proteins Palt, P20, P23, P24, several unidentified proteins, and did not contain Pwac, P23, and P22, 5) the yield of viable phage varied from 0.05 to 15% per cell. Thus, the entire morphogenesis of T4 phage can occur without accompanying replication of phage DNA.

摘要

噬菌体DNA在被噬菌体T4DtsLB3(基因42)感染的大肠杆菌B细胞中积累,此时不合成晚期蛋白质(在氯霉素存在的情况下)。然后(第二阶段),去除氯霉素,并用羟基脲抑制噬菌体DNA的进一步复制,同时将温度升至40摄氏度。使用M9培养基或含1%氨基酸的M9培养基;氯霉素的添加时间和羟基脲浓度也有所变化。还表明,在M9培养基中,在第二阶段主要合成早期蛋白质。在含氨基酸的培养基中,在第二阶段观察到以下情况:1)DNA合成完全被抑制,并且发生了DNA降解;2)早期和晚期蛋白质都有合成,其中晚期蛋白质占优势;3)发生了噬菌体尾部和衣壳元件的组装,但没有颈部和鞭毛,并且还发现了少量噬菌体颗粒;4)用氯仿裂解后在蔗糖密度梯度中分离出的衣壳含有蛋白质Palt、P20、P23、P24、几种未鉴定蛋白质,并且不含有Pwac、P23和P22;5)每个细胞中活噬菌体的产量从0.05%到15%不等。因此,T4噬菌体的整个形态发生过程可以在没有噬菌体DNA伴随复制的情况下发生。

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