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Lian-Qun Wang,De-Wu Liu,Wei Lan,Zun-Wen Lin,Pei-Xing Huang 한국조직공학과 재생의학회 2015 조직공학과 재생의학 Vol.12 No.4
Telomerase extends the proliferation and prevent replicative senescence in most somatic cells. Whether it has similar function in human epidermal stem cells remains to be determined. In this study, the human telomerase reverse transcriptase (hTERT) cytalytic subunit was introduced into epidermal stem cells derived from human fetal skins. The expression of hTERT mRNA, protein and telomerase activity in the transduced cells was observed by real time PCR (RT-PCR) techniques, western blot analysis and telomeric repeat amplification protocol enzyme-linked immunosorbent assay (TRAP-ELISA) assay, respectively. The proliferation of the transduced cells was examined. The results showed the introduction of hTERT into the epidermal stem cells upregulated the expression of the hTERT gene and protein. And also the hTERT-transduced epidermal stem cells exhibited significantly elevated telomerase activity and proliferation. These works establish the base of further gene modification, monoclone screening and cell differentiation mechanism. The ability to maintain the biological characteristics and proliferation of epidermal stem cells could have important applications in wound healing and skin tissue engineering.
( Qun Xia Li ),( Hong Lian Zhang ),( Yi Shan Lu ),( Jia Cai ),( Bei Wang ),( Ji Chang Jian ) 한국미생물 · 생명공학회 2015 Journal of microbiology and biotechnology Vol.25 No.3
Nocardia salmonicida is one of the main pathogens of fish nocardiosis. The purpose of this study was to build a loop-mediated isothermal amplification (LAMP) method for the rapid and sensitive detection of N. salmonicida. A set of four primers were designed from the 16S-23S rRNA intergenic spacer region of N. salmonicida, and conditions for LAMP were optimized as incubating all the reagents for 60 min at 64°C. LAMP products were judged with agar gel electrophoresis as well as with the naked eye after the addition of SYBR Green I. Results showed the sensitivity of the LAMP assay was 1.68 × 103 CFU/ml (16.8 CFU per reaction) and 10-fold higher than that of PCR. The LAMP method was also effectively applied to detect N. salmonicida in diseased fish samples, and it may potentially facilitate the surveillance and early diagnosis of fish nocardiosis.