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Updated: Jun 2, 2026
CAPRRESI: Chimera Assembly by Plasmid Recovery and Restriction Enzyme Site Insertion
Published on: June 25, 2017
[Construction of a shuttle plasmid encoding a chimeric gene of eglA p-Her2/neu ECD-IL-12]
Ying Liu1, Wen-qing Zhang, Hong Yu
1Laboratory of Pathogenic Biology; Medical College, Qingdao University, Qingdao 266071, China. shenghua005@163.com
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.
