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使用Bgl I、Bam HI、Hha I和Sma I限制性内切核酸酶对禽痘病毒进行限制性片段分析。

Restriction fragment analysis of fowlpox virus using Bgl I, Bam HI, Hha I and Sma I restriction endonucleases.

作者信息

Mishra S S, Mallick B B

机构信息

Indian Veterinary Research Institute, Izatnagar, India.

出版信息

Indian J Exp Biol. 1996 Oct;34(10):959-63.

PMID:9055646
Abstract

Fowlpox virus vaccine strain and two field isolates collected from out breaks of disease were purified from cell culture using sucrose gradient ultracentrifugation. The viral DNAs were digested with Bgl I, Bam HI, Hha I and Sma I restriction endonucleases and the fragment pattern was analysed on 0.7% agarose gel. Bgl I digestion produced 54 fragments of size ranging from 31.50 to 0.60 kb, having similar electrophoretic mobilities in both the vaccine strain and two field isolates. Only 9 well resolved and one unresolved or partially digested fragment were obtained after Bam HI digestion. A total number of 29 and 41 fragments were obtained with Hha I and Sma I respectively. Almost similar restriction fragment pattern was observed in vaccine strain and the field isolates. The total genomic size was calculated to be between 265.00 and 302.91 kb. The three viruses were found to be genetically similar.

摘要

禽痘病毒疫苗株以及从疾病暴发中收集的两株野外分离株,通过蔗糖梯度超速离心从细胞培养物中纯化得到。用Bgl I、Bam HI、Hha I和Sma I限制性内切酶消化病毒DNA,并在0.7%琼脂糖凝胶上分析片段模式。Bgl I消化产生了54个大小在31.50至0.60 kb之间的片段,在疫苗株和两株野外分离株中具有相似的电泳迁移率。Bam HI消化后仅获得9个分辨率良好的片段和1个未解析或部分消化的片段。Hha I和Sma I分别产生了29个和41个片段。在疫苗株和野外分离株中观察到几乎相似的限制性片段模式。计算得出总基因组大小在265.00至302.91 kb之间。发现这三种病毒在基因上相似。

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