Construction of Porcine Growth Hormone Eukaryotic Expression Vector and Its Transfection Mediated by Cationic Liposome in Mice

Abstract

The present study was designed to construct the eukaryotic expression vector for pGH mature peptide (mpGH) and to investigate its transfection mediated by cationic liposome (CLs) in COS-7 cells and mice. The cDNA of mpGH ORF was successfully cloned by reverse transcription-PCR (RT-PCR) using the adult pig pituitary gland RNA. The recombinant eukaryotic expression vector, VmpGH, was constructed by ligating the cDNA fragment to the vector VR1020. The successful construction was confirmed by restriction enzyme digestion, and the expression of mpGH was confirmed by RT-PCR, immunofluorescence analyses (IFA), and ELISA in COS-7 cells. The VmpGH and VR1020 plasmids were entrapped with CLs, and four experimental groups of male Kunming mice were administrated with VmpGH / lipoplex or naked VmpGH plasmids at two dosages (0.5 and 1.0 mg/kg), while the mice injected with VR1020-lipoplex at the dosage of 0.5 mg/kg body weight (BW) were used as control. The BWs of the mice administrated with VmpGH-lipoplex at both dosages were significantly higher than not only those of the control (P < 0.01) but also those of mice injected with naked plasmids (P < 0.01), from 30 to 60 days post-transfection. The transcription of VmpGH was detected by RT-PCR in six tissues, including the liver, kidney, spleen, heart, muscle, and blood, of the mice injected with VmpGH-lipoplex, but not in the same tissues of control mice. Furthermore, the mice injected with VmpGH-lipoplex showed higher plasma GH contents than the control mice (P < 0.05), although their IgG contents did not show much difference. Our study demonstrates that the VmpGH plasmids' transfection mediated by CLs can significantly promote the growth of mice, which may be used to improve the livestock production.

Keywords:

• Cationic liposome (CLs)
• Growth-promoting
• Porcine growth hormone (pGH)
• Transfection

Acknowledgments

Chenchen Peng and Lihong Wang contributed equally to this work

Notes

^a,b Indicates significant differences (p ≤ 0.01).

Note: Group I indicates mice injected with VmpGH/lipoplex at a dosage of 0.5 mg/kg BW and Group II injected with VmpGH/lipoplex at a dosage of 1.0 mg/kg; Group III indicates the mice injected with naked VmpGH plasmid at a dosage of 0.5 mg/kg BW and Group IV at 1.0 mg/kg BW; Group V indicates the mice injected with VR1020/lipoplex at a dosage of 0.5 mg/kg BW.

^A,B,C,D,E Indicates significant differences (p ≤ 0.01).

^a,b,c,d,e Indicates specific differences (p ≤ 0.05) in rows.

^a,b Indicates the compared groups are different at p < 0.05.

^a,b Indicates the mean difference of the compared groups is significant at the 0.05 level (p < 0.05).