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限制性内切酶克隆方法，以评估<em>在体外</em> HIV-1 C亚型的Gag-MJ4嵌合病毒的复制能力
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Immunology and Infection
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JoVE Journal Immunology and Infection
A Restriction Enzyme Based Cloning Method to Assess the In vitro Replication Capacity of HIV-1 Subtype C Gag-MJ4 Chimeric Viruses
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限制性内切酶克隆方法，以评估<em>在体外</em> HIV-1 C亚型的Gag-MJ4嵌合病毒的复制能力

DOI:
10.3791/51506-v

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14:23 min

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August 31, 2014

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, ,
1Emory Vaccine Center at Yerkes National Primate Research Center,Emory University, 2Department of Pathology and Laboratory Medicine,Emory University

Chapters

  • 00:05Title
  • 01:53Amplification of the HIV-1 Gag Gene from Infected, Frozen Plasma
  • 03:29Cloning Amplified Gag Genes into the MJ4, Subtype C, Infectious Molecular Clone
  • 06:09In vitro Replication of Gag-MJ4 Chimeras in GXR25 (CEM-CCR5-GFP) Cells
  • 10:06Analysis of Reverse Transcriptase in Cell Culture Supernatants
  • 12:11Results: A Restriction Enzyme Based Cloning Method to Assess the In vitro Replication Capacity of HIV-1 Subtype C Gag-MJ4 Chimeric Viruses
  • 13:44Conclusion

Summary

Automatic Translation

Automatic Translation

HIV-1 pathogenesis is defined by both viral characteristics and host genetic factors. Here we describe a robust method that allows for reproducible measurements to assess the impact of the gag gene sequence variation on the in vitro replication capacity of the virus.

Tags

Keywords: HIV-1Subtype CGagReplication CapacityRestriction Enzyme CloningMJ4ZambiansCellular Immune PressureHLAViral LoadCD4+ T Cell Decline

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