Identification, Characterization, and Expression Analysis of Immunoglobulin Genes from Antarctic Fish by PCR Methods

Alessia Ametrano; Maria Rosaria Coscia

doi:10.1007/978-1-0716-2313-8_20

Identification, Characterization, and Expression Analysis of Immunoglobulin Genes from Antarctic Fish by PCR Methods

Methods Mol Biol. 2022:2498:351-362. doi: 10.1007/978-1-0716-2313-8_20.

Authors

Alessia Ametrano^{1

2}, Maria Rosaria Coscia³

Affiliations

¹ Institute of Biochemistry and Cell Biology-National Research Council of Italy, Naples, Italy.
² Department of Environmental, Biological and Pharmaceutical Sciences and Technologies, University of Campania Luigi Vanvitelli, Caserta, Italy.
³ Institute of Biochemistry and Cell Biology-National Research Council of Italy, Naples, Italy. [email protected].

PMID: 35727556
DOI: 10.1007/978-1-0716-2313-8_20

Abstract

We describe different applications of the polymerase chain reaction (PCR) method to amplify up to 2.5-kb DNA regions of the immunoglobulin genes from Antarctic teleost fishes and to evaluate their expression in different tissues. To enhance amplification of poorly expressed transcripts or difficult to amplify, for example, products generated by 5' or 3' RACE, we have set up optimal primer annealing conditions based on a three-step cycle strategy.The protocols reported here along with materials required and general tips are modifications of conventional PCR.

Keywords: 5′ and 3′ RACE; Antarctic teleost; Cold-adapted gene; IgH constant region gene; PCR; qPCR.

MeSH terms

Animals
Antarctic Regions
Base Sequence
Fishes / genetics
Genes, Immunoglobulin*
Immunoglobulin Heavy Chains* / genetics
Polymerase Chain Reaction

Substances

Immunoglobulin Heavy Chains