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Construction method and application of eukaryotic expression linear vector based on avian reticuloendotheliosis virus LTR

A linearized vector, avian reticuloendothelial technology, applied in biochemical equipment and methods, applications, botanical equipment and methods, etc., can solve the problems of cumbersome cloning process and low efficiency, and achieve the goal of improving efficiency and simplifying the cloning process. Effect

Inactive Publication Date: 2015-06-17
YANGZHOU UNIV
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Problems solved by technology

But sometimes the cloning process is cumbersome and inefficient due to the inability to find a suitable restriction site

Method used

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  • Construction method and application of eukaryotic expression linear vector based on avian reticuloendotheliosis virus LTR
  • Construction method and application of eukaryotic expression linear vector based on avian reticuloendotheliosis virus LTR
  • Construction method and application of eukaryotic expression linear vector based on avian reticuloendotheliosis virus LTR

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Embodiment

[0026] 1) Design and amplify the linearized vector containing REV virus LTR and primers for EGFP fragments: the upstream primer for amplifying the linearized vector containing REV virus LTR is located at position 7783-7811 of the proviral genome of the SNV strain; amplify the linearized vector containing REV virus LTR The downstream primer of the eGFP vector is located at position 974-1002 of the proviral genome of the SNV strain; in addition to the 18 bases specific to the eGFP fragment, the upstream primer for amplifying the EGFP fragment also contains 15 conserved and amplified REV virus-containing LTRs Bases that are complementary to the downstream primers of the linearized vector; in addition to the 18 bases N specific to the EGFP fragment, the downstream primers for amplifying the eGFP fragment also contain 15 conserved and upstream primers for the linearized vector that amplifies the LTR of the REV virus complementary bases. See attached table 1 for specific primer sequ...

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Abstract

The invention relates to a construction method and application of a eukaryotic expression linear vector based on an avian reticuloendotheliosis virus LTR. The method comprises the following steps: performing PCR amplification utilizing primers of which the sequences are shown as SEQ ID NO.1 and 2, taking an SNV strain as a template, thereby obtaining the REV virus LTR linear vector. The method disclosed by the invention can be applied to research of duplication, translation and pathogenicity of REV or other retroviruses and other mechanisms; and moreover, rapid cloning and high-efficiency expression of exogenous genes can be realized.

Description

technical field [0001] The invention relates to the construction and application of a eukaryotic expression vector based on avian reticuloendotheliosis virus LTR. The invention can not only be used to study the replication, translation, pathogenicity and other mechanisms of REV or other retroviruses, but also can realize rapid cloning and high-efficiency expression of foreign genes. Background technique [0002] Avian reticuloendotheliosis virus (REV) mainly causes a group of syndromes characterized by reticuloendothelial cell hyperplasia, including acute reticulosis, dwarf syndrome and chronic neoplastic hyperplasia of lymphoid tissue and other tissues disease. REV-infected chickens tend to grow stunted, and the elimination rate and mortality increase; and cause immunosuppression of infected chickens, interfere with the immune effect of other poultry disease vaccines, resulting in immune failure. The REV proviral genome structure has a complete retroviral genome structure...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/867C12N15/66
Inventor 叶建强田晓彦范中雷秦爱建邵红霞
Owner YANGZHOU UNIV
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