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Expression, Isolation, and Purification of Soluble and Insoluble Biotinylated Proteins for Nerve Tissue Regeneration
Published on: January 22, 2014
Cloning and expression of three peptide-linked beta2-microglobulin molecules in Escherichia coli with an isocaudamer
Fanyan Meng1, Chuanlai Shen, Yong He
1Key Laboratory of Ministry of Education for Developmental Genes and Human Disease, Department of Genetics and Developmental Biology, Southeast University Medical School, Nanjing 21009, People's Republic of China.
Abstract:
The application of the peptide-linked beta2-microglobulin (beta2m) strategy is limited in some cases due to the incompatibility between the sequences of the peptides and the restriction sites of the plasmid vectors. An isocaudamer technique was adapted to overcome this restriction. Three peptide-linked beta2m genes, HBc(18-27)-hbeta2m gene, OVA(257-264)-mbeta2m gene and HER2/neu(369-377)-mbeta2m gene, were inserted into the pET28a vectors with this technique. The corresponding proteins were expressed in Escherichia coli with yields of over 50 mg/l culture and purities of over 80%. This strategy facilitates the construction of peptide-linked beta2m molecules and will simplify the preparation of major histocompatibility complex-peptide complexes.
