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Updated: Jul 5, 2026

Functional Cloning Using a Xenopus Oocyte Expression System

Functional Cloning Using a Xenopus Oocyte Expression System

Published on: January 30, 2016

cDNA expression cloning in mammalian cells.

¹National Institute of Mental Health, Bethesda, Maryland, USA.

Current Protocols in Neuroscience

|April 23, 2008

Summary

This summary is machine-generated.

This study details protocols for expression cloning in mammalian cells using various transfection methods and screening assays. Establishing a robust bioassay for the gene product is crucial before library construction and cloning.

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Published on: February 9, 2015

Area of Science:

Molecular Biology
Genetics
Cell Biology

Background:

Expression cloning in mammalian cells is essential for functional genomics.
Screening cDNA libraries requires efficient methods for identifying expressed genes.

Purpose of the Study:

To provide detailed protocols for expression cloning in mammalian cells.
To outline methods for preparing and screening cDNA libraries.

Main Methods:

Utilizing calcium phosphate or liposome-mediated transfection for mammalian cell expression.
Employing virus infection combined with liposome-mediated transfection.
Preparing cDNA pools from library-transformed E. coli.
Screening expression using autoradiography, direct visualization, functional assays (scintillation counting), or filter-based binding assays.

Main Results:

Successful implementation of various transfection and screening techniques.
Demonstration of methods for detecting and quantifying gene expression.
Identification of critical steps in the cDNA cloning process.

Conclusions:

The establishment of a specific bioassay for the gene product is paramount.
Effective screening protocols are key to successful expression cloning projects.
These protocols facilitate the identification of functional genes within mammalian systems.