Cloning and Overexpression

Cloning and subcloning

CBI uses standard restriction enzyme, PCR, and ligation technology to clone or subclone known genes. RT-PCR can be applied to clone cDNAs of interest from supplied RNA, or RNA can be purified by CBI. 5’-RACE-PCR can be performed to identify and clone the 5’ end of a cDNA. 3’RACE can be applied if the 3’ end is unknown. Genes can be recloned to construct full length clones.

Overexpression

CBI will design your protein for overexpression in E. coli, yeast, or baculoviral systems. Proteins can be designed with “his-tags”, “GST-tags”, or other system of your choice. CBI recommends a regulatable expression system to provide optimum protein yields, but will work with the vector system of your selection. Overexpressed proteins are analyzed by SDS-PAGE, purified using the affinity tags or other chromatographic step, and verified by N-terminal sequence analysis, if requested. Standard proteins are provided at ~85% purity. Additional purification steps are available at your discretion.

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Please call 800-735-9224 or e-mail for additional information or a quotation.