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The Chinese Journal of Dental Research

Year 2001
Volume 4 , Issue 3

Pages: 32 - 36

Establishment of Rat Tongue Model for Plasmid DNA Expression in Vivo

ZeBing Zhang, PhD/WenTao Gao, PhD/Jie Ouyang, PhD

Objective: To develop rat tongue model for plasmid DNA expression in vivo. Methods: The expression plasmid DNA, which contained either human muscle-specific promoter or SV40 promoter directing the transcription of E. coli LacZ reporter gene, was injected directly into Wistar rat tongue. PCR technique, X-gal histochemical staining, and Southern blot hybridization were used to analyze the reporter gene expression in vivo. Results: E. coli LacZ gene sequence was found in rat tongue injected by PCR with the plasmid DNA, which demonstrated the capacity of rat tongue myofibers to uptake foreign DNA. The expression of reporter genes was correlated with the amount of injected plasmid DNA and the incubation time in tongue muscle. Β-galactosidase activity was detected 24 hours after injection by X-gal staining, but maximal expression level was observed one week later, and reporter gene expression could even be detected at the 2-month time point. Co-injection of the 2 expression plasmids resulted in co-expression in the same striated myofibers, as well as in individual expression in different myofibers. The Southern blot analysis showed the injected plasmid DNA existing outside the genome of rat muscle cell. No integration was detected. Conclusion: These results show that direct plasmid DNA injection into tongue muscle is a simple and efficient approach to transfer foreign genes into the striated muscle cells of tongue, and that tongue is a good model for analyzing exogenous gene expression.



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