Author(s)

Lin Qiu, Chunbao Guo

Laboratory of Surgery, Children’s Hospital of Chongqing Medical University, Chongqing, China.
Laboratory of Surgery, Children’s Hospital of Chongqing Medical University, Chongqing, China;.

In this study, we cloned human KGF (hKGF) genes using RT-PCR techniques and developed a eukaryotic expression plasmid vector capable of directing the expression of functional hKGF. Monolayer culture of human embryo lung fibro-blast (HLF) was used for isolation of total RNA. Then the total RNA was purified and reverse- transcribed into cDNA using an oligo (dT) primer. A full PCR fragment for hKGF was generated and cloned. Restriction digestion and nucleo-tide sequence analysis validated the complete hKGF transcription. The hKGF cDNA fragment was inserted into pEGFP-C2 vector by means of recombinant DNA technology and verified by restriction analysis and sequencing. We have constructed pEGFP-C2-hKGF encoding the green fluorescent protein (GFP). Furthermore, hKGF had the effect on AEC II proliferation. These results suggest that the potential appli-cation of a hKGF plasmid of gene expression should be useful for sustained AEC proliferation, and its in vivo efficacy needs to be validated. Keywords:

Human Embryo Lung Fibroblast; Gene Clone; Reverse Transcriptage Polymerase Chain Reaction; Eukaryotic Expression Vector

Qiu, L. and Guo, C. (2010) A Plasmid vector encoding functional human keratinocyte growth factor gene in vitro—Functional human KGF gene expression in vitro. Journal of Biophysical Chemistry, 1, 64-71. doi: 10.4236/jbpc.2010.11008.