Abstract
Collagen에 의해 유도되는 류마티스 관절염을 저해하는 효과가 있는 인간 histone 단백질 Hl.5 (hHl.5)를 산화스트레스 유도성 SWPA2 프로모터에 연결하여 형질전환 담배(Nicotiana tabacum L. cv Bright Yellow-2) 배양세포주를 개발하였다. hH1.5 유전자는 Agrobacterium 매개 형질전환 방법으로 담배 BY-2 배양세포에 도입되었다. 형질전환 캘러스는 150mg/L kanamycin과 300mg/L claforan이 포함된 변형된 MS 선발배지에서 선발하여, PCR분석으로 hHl.5 유전자의 도입을 확인하였다. 형질전환 현탁배양세포에서 hH1.5 단백질의 발현은 northern 분석과 Western 분석으로 확인하였는데, 담배배양세포에서 재조합hHl.5 단백질 (42 kDa)은 인간의 것 (32 kDa)과는 다른 크기의 단백질이 확인되었다. 금후 재조합 hH1.5 단백질의 자세한 특성규명이 요구된다. Transgenic tobacco (Nicotiana tabacum L. cv Bright Yellow-2) cell lines expressing a human histone H1.5 (referred to as hH1.5), which suppress collagen-induced rheumatoid arthritis, were developed under the oxidative stress-inducible peroxidase (SWPA2) promoter. Tobacco BY-2 cells were transformed by Agrobacterium-mediated method. The kanamycin-resistant calli were selected on the modified MS medium containing 150mg/L kanamycin and 300mg/L claforan. Transgenic cell lines were confirmed by PCR and northern blot analysis. Recombinant hH1.5 (rhH1.5) protein (42 kDa) was also detected by Western blot analysis, showing a different molecular weight of human hH1.5 (32 kDa). These results suggested that a hH1.5 gene was properly introduced in tobacco cultured cells under the control of SWPA2 promoter. The further characterization of rhH1.5 protein remains to be studied.
Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
