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Building method and applications of J avian leukosis virus subgroup LTR based linearized eukaryotic expression vectors

A technology of avian leukosis virus and eukaryotic expression vector, applied in the field of construction and application of linearized eukaryotic expression vector based on J subgroup avian leukosis virus LTR, can solve problems such as low efficiency and cumbersome cloning process, and achieve improved efficiency, The effect of simplifying the cloning process

Inactive Publication Date: 2015-06-24
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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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  • Building method and applications of J avian leukosis virus subgroup LTR based linearized eukaryotic expression vectors
  • Building method and applications of J avian leukosis virus subgroup LTR based linearized eukaryotic expression vectors
  • Building method and applications of J avian leukosis virus subgroup LTR based linearized eukaryotic expression vectors

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Experimental program
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Embodiment

[0026] 1) Design and amplify the linearized vector containing the ALV-J virus LTR and primers for the eGFP fragment: the upstream primer of the linearized vector containing the ALV-J virus LTR is located 28 bases after the stop codon of the ALV-J env gene; The downstream primer of the linearized vector containing the ALV-J virus LTR is located 27 bases before the start codon of the ALV-Jgag gene; the upstream primer for amplifying the eGFP fragment contains 18 bases specific to the eGFP fragment. 15 conservative bases complementary to the downstream primers of the linearized vector containing the ALV-J virus LTR; the downstream primers for amplifying the eGFP fragment contain 15 conservative bases in addition to the eGFP fragment-specific 18 bases The base complementary to the upstream primer of the linearized vector containing the ALV-J virus LTR. The specific primer sequences are shown in Table 1, synthesized by Life Technologies Shanghai Thermo Fisher Scientific.

[0027] 2) ...

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Abstract

The invention relates to a building method and applications of J avian leukosis virus subgroup LTR based linearized eukaryotic expression vectors. According to the invention, by taking sequences shown in SEQ ID NO.1 and SEQ ID NO.2 as primers and taking a JSNT strain as a template, an ALV-J virus LTR linearized vector is obtained through PCR amplification. The building method and applications of J avian leukosis virus subgroup LTR based linearized eukaryotic expression vectors disclosed by the invention not only can be used for researching the replication, translation and pathogenic mechanisms and the like of ALV-J or other retroviruses, and also can realize the rapid cloning and efficient expression of exogenous genes.

Description

Technical field [0001] The invention relates to the construction and application of a linearized eukaryotic expression vector based on J subgroup avian leukemia virus LTR. This invention can not only be used to study the mechanisms of ALV-J or other retroviruses such as replication, translation, and pathogenicity, but also can realize rapid cloning and high-efficiency expression of foreign genes. Background technique [0002] According to viral envelope protein antigenicity, interference neutralization test and host range, avian leukemia virus (ALV) can be divided into 10 subgroups (A-J). Unlike other ALVs, ALV-J infection mainly causes malignant proliferation of hematopoietic cells, immunosuppression, and induces tumors such as myeloid tumors and hemangioma. The ALV-J proviral genome structure has a complete retroviral genome structure, including the 5'LTR-gag-pol-env-LTR3' basic structure. Among them, LTR is a non-coding long terminal repeat sequence, which is closely related...

Claims

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Application Information

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IPC IPC(8): C12N15/867
Inventor 叶建强范中雷田晓彦秦爱建邵红霞
Owner YANGZHOU UNIV