Introduction

After cloning a gene of interest, many researchers wish to analyze its characteristics by overexpression analysis or by introduction of mutated forms of the gene of interest into various cell types. In the analysis of insulin-stimulated glucose transport, the most appropriate cell systems are striated muscle and adipocytes (1). However, the introduction of DNA or genes of interest into these insulin-responsive tissues by standard transfection protocols such as calcium phosphate, DEAE-dextran, and liposome-mediated transfection are very inefficient. Furthermore, although transgenes can be expressed in muscle using adenovirus infection systems, this is difficult to accomplish in adipocytes and is substantially more labor intensive. The production of recombinant adenoviruses to use in infection of insulin-responsive tissues can take several months and requires very high titers of adenovirus. Therefore, we have recently established electroporation conditions that consistently provide at least 50% transfection efficiency for cultured differentiated 3T3-L1 adipocytes (2). Although the electroporation is not 100% efficient, it provides an easy and fast method to introduce DNA into adipocytes. Using 600 ig of CMV-LacZ plasmid DNA, we consistently obtain an electroporation efficiency of 50-80% (see Fig. 1).

Diabetes 2

Diabetes 2

Diabetes is a disease that affects the way your body uses food. Normally, your body converts sugars, starches and other foods into a form of sugar called glucose. Your body uses glucose for fuel. The cells receive the glucose through the bloodstream. They then use insulin a hormone made by the pancreas to absorb the glucose, convert it into energy, and either use it or store it for later use. Learn more...

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