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Stability and Structure of Bat Major Histocompatibility Complex Class I with Heterologous β2-Microglobulin
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Immunologia e infezioni
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JoVE Journal Immunologia e infezioni
Stability and Structure of Bat Major Histocompatibility Complex Class I with Heterologous β2-Microglobulin

Stability and Structure of Bat Major Histocompatibility Complex Class I with Heterologous β2-Microglobulin

DOI:
10.3791/61462-v

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11:17 min

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March 10, 2021

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, , , , , , , , , ,
1School of Laboratory Medicine and Life Sciences,Wenzhou Medical University, 2NHC Key Laboratory of Biosafety, National Institute for Viral Disease Control and Prevention,Chinese Center for Disease Control and Prevention, 3Faculty of Health Sciences,University of Macau, 4CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology,Chinese Academy of Sciences, 5Savaid Medical School,University of Chinese Academy of Sciences

Capitoli

  • 00:04Introduction
  • 00:54E. coli Transformation
  • 01:46Culture Inoculation
  • 02:59Bacteria Harvest
  • 03:28Inclusion Body Purification
  • 04:27MHC Complex Refolding
  • 04:53MHC H Chain and Beta2m Injection and Dilution
  • 05:42Refolded Protein Concentration Determination
  • 06:51Crystallization, Data Collection, and Processing
  • 07:41Structure Determination and Analyses
  • 08:33Results: Representative Bat MHC I Beta2-Microglobulin Stability and Structure Analysis
  • 10:21Conclusion

Summary

Traduzione automatica

Traduzione automatica

The protocol describes experimental methods to obtain stable major histocompatibility complex (MHC) class I through potential β2-microglobulin (β2m) substitutions from different species. The structural comparison of MHC I stabilized by homologous and heterologous β2m were investigated.

Tags

Bat MHC Class IHeterologous β2-microglobulinHybrid MHC Class IE. Coli TransformationRecombinant Protein ExpressionIPTG InductionBacterial Cell PelletPBS

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