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Construction and Evaluation of Novel Plasmid DNA Encoding Somatostatin Fused to a Tissue Plasminogen Activator (tPA) in Mice

Construction and Evaluation of Novel Plasmid DNA Encoding Somatostatin Fused to a Tissue Plasminogen Activator (tPA) in Mice

Fangxiao Dong1,2, Xuan Luo1,2, Delu Yuan1,2, Aixin Liang1,2* and Liguo Yang1,2*

1Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, (Huazhong Agricultural University), Ministry of Education, Wuhan 430070, PR China
2College of Animal Science and Technology &College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, PR China

Aixin Liang and Liguo Yang contributed equally in this article.

*      Corresponding author: yangliguo889555@163.com

 

Fig. 1.

Construction of plasmid pEGS/2SS-tPA. The sequence of tissue plasminogen activator signal peptide (tPA) was inserted into previous somatostatin DNA vaccine (pEGS/2SS). A, the plasmid profile of pEGS/2SS-tPA; B, The sequencing analysis of pEGS/2SS-tPA.

Fig. 2.

Expression and distribution of green fluorescent in MCF-7 cells at 24 h and 48 h after transfection. Scale bar = 200μm.

Fig. 3.

Relative expression of SS mRNA by real-time PCR. The Ct value of SS was normalized to that of GAPDH, and the SS expression level in cells transfected with pEGS/2SS-tPA or pEGS/2SS was calculated relative to cells transfected with pEGFP-N1. **P<0.01.

Fig. 4.

S/2SS fusion protein detection in cell supernatants by Western blot. Lane 1, cells transfected with pEGS/2SS-tPA; Lane 2, cells transfected with pEGS/2SS; Lane 3, cells transfected with negative control pEGFP-N1. GAPDH protein was selected as the standardized correction of S/2SS fusion protein expression quantity.

Fig. 5.

Relative expression of PRLR, JAK2, STAT5a and DGAT1 by Real-time PCR. The Ct value of SS was normalized to that of GAPDH, and the expression level of targets genes in cells transfected with pEGS/2SS-tPA or pEGS/2SS was calculated relative to cells transfected with pEGFP-N1. *P<0.05.

Fig. 6.

Serum GH concentration (ng/mL) and serum PRL concentration (ng/mL) in T1, T2 and C groups vaccinated with pEGS/2SS-tPA, pEGS/2SS and pEGFP-N1 at 8 week after immunization. *P<0.05 and **P<0.01.

Pakistan Journal of Zoology

October

Vol. 50, Iss. 5, Pages 1601-1998

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