KSBB Journal

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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Analysis of UreB Protein Synthesis from Transgenic Lily Pollen

형질전환 백합화분을 이용한 UreB단백질의 발현분석

  • 박희성 (대구가톨릭대학교 생명자원학부) ;
  • 박인혜 (대구가톨릭대학교 생명자원학부)
  • Published : 2002.12.01
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Abstract

In an attempt to produce recombinant proteins using the pollen enriched in some plant species, a 1.7 kb DNA encoding urease subunit B (UreB) amplified by PCR from Helicobacter pylori urease gene cluster in pH808 plasmid was cloned to be expressed under CaMV35S promoter in lily (Lilium longiflorum) pollen tubes elongated in vitro. Lily pollen at early germinating stage was transformed with the ureB DNA using Agrobacterium via vacuum infiltration and, incubated for a full pollen tube growth 16 - 24 h in the dark in the presence of kanamycin. DNA integration and expression in the transgenic pollen were analyzed by the standard molecular techniques and the results suggest that the pollen in vitro may be employed as a protein factory in a disposable fashion.

풍부한 식물의 화분을 이용하여 재조합단백질의 생산연구를 위하여 UreB 단백질 정보를 지닌 1.7 kb DNA를 Helicobacter pylori urease gene cluster를 지니는 pH808로보터 PCR을 통하여 증폭하고 이를 CaMV35S promoter에 연결하여 백합(Lilium longiflorum)화분내로 도입하고 기내배양을 실시하였다. 발아초기의 화분을 Agrobacterium과 함께 진공침윤시켜 ureB DNA를 형질전환시키고 kanamycin을 지니는 화분배지에서 16-24시간 배앵하여 완전한 화분관신장을 이루도록 하였다. 이들 형질전환화분의 유전자도입 및 발현을 분석하였으며 그 결과 기내배양하는 하분을 일회성의 단백질공장으로 이용할 수 있다는 가능성을 제시하였다.

Keywords

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