DNA packaging and developmental intermediates of a broad host range Vibrio vulnificus bacteriophage 71A-6.
DNA packaging and developmental intermediates of a broad host range Vibrio vulnificus bacteriophage 71A-6.
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广泛宿主范围创伤弧菌噬菌体 71A-6 的 DNA 包装和发育中间体。
DOI：
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发表时间：
2001
影响因子：
3.3
通讯作者：
C. Cerniglia
中科院分区：
文献类型：
--
作者：
S. Khan;A. Khan;M. Nawaz;A. DePaola;A. Andrews;C. Cerniglia
采用超速离心法分离了创伤弧菌噬菌体71a-6杆状双链DNA痘病毒衣壳组装和DNA包装途径中的结构中间体，并用电子显微镜、SDS-PAGE和脉冲场凝胶电泳法对其进行了研究。噬菌体71a-6在发育过程中合成棒状衣壳(平均长度为200+/-8 nm，平均宽度为47+/-3 nm，n=50)。几种可能有助于衣壳组装和成熟的宿主蛋白以球形结构附着在这些衣壳上。还分离到一个衣壳-DNA或DNA包装复合体，由成熟衣壳、DNA和一个42.5 kDa的蛋白质组成。DNA包装前后衣壳长度增加，宽度减小(平均长度220+/-8 nm；平均宽度45+/-3 nm n=50)。衣壳部分含有约12个噬菌体结构蛋白和8个宿主蛋白。初步鉴定了至少3种蛋白质：38.5 kDa的主要衣壳蛋白、35.2 kDa的尾蛋白和42.5 kDa的包装启动子或终止子蛋白。经脉冲场凝胶电泳法测定，71a-6噬菌体基因组大小为143.0 kb。所有成熟噬菌体蛋白的总质量仅占噬菌体基因组编码量的14.0%。
The structural intermediates in the capsid assembly and DNA packaging pathway of Vibrio vulnificus bacteriophage 71A-6, a rod-shaped double-stranded DNA podovirus, were isolated by ultracentrifugation and studied by electron microscopy, SDS-PAGE and pulsed-field gel electrophoretic analysis. Bacteriophage 71A-6 synthesized rod-shaped capsids (mean length=200+/-8 nm; mean width=47+/-3 nm n=50) during its development. Several host proteins that probably help in the assembly and maturation of the capsids were attached to these capsids as spherical structures. A capsid-DNA or DNA packaging complex that consisted of the mature capsids, DNA and a 42.5-kDa protein was also isolated. The size of the capsids increased in length and decreased in width (mean length=220+/-8 nm; mean width=45+/-3 nm n=50) either during or after the DNA packaging. The capsid fractions contained about 12 phage structural proteins and eight host proteins. At least three proteins were tentatively identified: a 38.5-kDa major capsid protein, a 35.2-kDa tail protein and 42.5-kDa packaging initiator or terminator protein. The size of the bacteriophage 71A-6 genome was determined to be 143.0-kb by pulsed-field gel electrophoresis. The total mass of all the mature phage proteins corresponded to only 14.0% of the coding capacity of phage genome.
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DOI：
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发表时间：
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期刊：
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影响因子：
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