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JoVE Journal

Immunology and Infection

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

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11:17 min

March 10th, 2021

March 10th, 2021

6,161 Views

0:04

Introduction

0:54

E. coli Transformation

1:46

Culture Inoculation

2:59

Bacteria Harvest

3:28

Inclusion Body Purification

4:27

MHC Complex Refolding

4:53

MHC H Chain and Beta2m Injection and Dilution

5:42

Refolded Protein Concentration Determination

6:51

Crystallization, Data Collection, and Processing

7:41

Structure Determination and Analyses

8:33

Results: Representative Bat MHC I Beta2-Microglobulin Stability and Structure Analysis

10:21

Conclusion

Transcript

In this study, we used Bat MHC Class I as a model to summarize the methodology, and to evaluate the feasibility of a hybrid MHC class I complexed with heterologous beta-2 microglobulin. Previous studies have shown that the mammalian B2M substituti

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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.

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