Construction and purification of ANK gene deleted recombinant goatpox virus
Construction and purification of ANK gene deleted recombinant goatpox virus
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ANK基因缺失重组羊痘病毒的构建及纯化
DOI：
10.1007/s13337-020-00620-z
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发表时间：
2020-08
期刊：
影响因子：
--
通讯作者：
Youwen Li
中科院分区：
文献类型：
--
作者：
Xueping Zhang;Jianjun Tong;· Tuohetiniyazi Milikaimu;Chuanchuan He;Wei Wang;Youwen Li
羊痘病毒（SPPV）和山羊痘病毒（GTPV）是两种具有宿主特异性的病原体。先前的研究假设锚蛋白（ANK）家族可能在确定SPPV和GTPV的宿主范围方面发挥重要作用。为了验证ANK蛋白的功能，生成并纯化ANK基因缺失的GTPV至关重要。本研究以GFP基因作为报告基因，通过融合PCR将ANK基因的两个同源臂连接起来。将PCR产物插入PET42b中，生成ANK基因缺失的转移载体，并转染至GTPV感染的睾丸原代细胞中。 rGTPV 被鉴定为绿色荧光阳性并经过适当纯化。结果表明，GFP基因与ANK基因的两个同源臂相连。将该序列插入PET42b中以形成ANK缺失的转移载体。通过将载体和GTPV转移到细胞中，成功地产生了ANK缺失的rGTPV。本研究中纯化并鉴定了 ANK 缺失的 rGTPV。该研究成功生成了ANK缺失的rGTPV。它克服了未来研究ANK基因功能的技术障碍。
Sheeppox virus (SPPV) and goatpox virus (GTPV) are two pathogens of host specificity. Previous studies have hypothesized that ankyrin (ANK) family may play an important role in determining host range of SPPV and GTPV. In order to verify the function of ANK proteins, it is critical to generate and purify the ANK gene deleted GTPV. In this study, the GFP gene as a reporter gene was connected with two homologous arms of ANK gene by fusion PCR. The ANK gene deleted transfer vectors were generated by inserting the PCR products into PET42b, and were transfected into testicular primary cells which were infected by GTPV. The rGTPV were identified as green fluorescence positive and properly purified. The results showed that GFP gene and two homologous arms of ANK gene were connected. The sequence was inserted in PET42b to form ANK deleted transfer vector. ANK deleted rGTPV was generated successfully by transferring vector and GTPV in cells. The ANK deleted rGTPV was purified and identified in this study. The study successfully generated the ANK deleted rGTPV. It overcomes the technical barrier for future studies about the function of ANK genes.
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DOI：
10.1038/nrmicro1099
发表时间：
2005-03
期刊：
Nature reviews. Microbiology
影响因子：
--
作者：
通讯作者：
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影响因子：
9.7
作者：
Chin, Jiah Shin;Chooi, Wai Hon;Chew, Sing Yian
通讯作者：
Chew, Sing Yian
影响因子：
4.4
作者：
Offerman K;Carulei O;van der Walt AP;Douglass N;Williamson AL
通讯作者：
Williamson AL
影响因子：
2.2
作者：
Oguzoglu, T. C.;Alkan, F.;Burgu, I.
通讯作者：
Burgu, I.
影响因子：
4.3
作者：
Adedeji, Adeyinka Jeremy;Moeller, Janika;Hoffmann, Bernd
通讯作者：
Hoffmann, Bernd