[Construction of a shuttle plasmid encoding a chimeric gene of eglA p-Her2/neu ECD-IL-12].

AIM: To construct a shuttle plasmid encoding a chimeric gene of Clostridium saccharobutylicum eglA promoter(eglA p)-extracellular domain of human epidermal growth factor receptors 2(hHer2/neu ECD)-human Interleukin-12(rhIL-12), pIMP1 eglA p-hHer 2/neu ECD-rhIL-12. METHODS: The hHer2/neu ECD and the eglA p was amplified from the corresponding template: pcDNA3.1 hHer2/neu and 55 bp fragment in eglA p by PCR. pcDNA6 eglA p-hHer2/neu ECD-rhIL-12 was prepared by inserting the hHer2/neu ECD and eglAp fragment into the plasmid, pcDNA6 rhIL-12. Shuttle plasmid pIMP1 eglA p-hHer2/neu ECD-rhIL-12 was acquired by using in-fusion technique. subsequently, the recombinant shuttle plasmid was identified. RESULTS: All the fragments of hHer2/neu ECD , eglA p and eglAp-Her2/neu ECD-rhIL-12 were amplified, and the corresponding plasmids were prepared rightly. The shuttle plasmid of pIMP1 eglAp-hHer2/neu ECD-rhIL-12 was successfully const- ructed. No error was found both in the sequence and ORF of the acquired chimeric gene. CONCLUSIONS: A shuttle plasmid encoding a chimeric gene of Clostridium saccharobut- ylicum eglAp-hHer2/neu ECD-rhIL-12(pIMP1 eglA p-hHer2/neu ECD-rhIL- 12) was success- fully constructed.