• 제목/요약/키워드: actinidin gene

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Isolation of Cysteine Protease Actinidin Gene from Chinese Wild Kiwifruit and its Expression in Escherichia coli

  • Lee, Nam-Keun;Hahm, Young-Tae
    • Food Science and Biotechnology
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    • 제16권2호
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    • pp.294-298
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    • 2007
  • The actinidin (EC 3.4.22. 14) found in kiwifruit is a cysteine protease. In order to obtain the actinidin gene from the Chinese wild kiwifruit, primers were designed on the basis of the actinidin gene of Actinidia deliciosa, the New Zealand kiwifruit. The 1.2 kb DNA fragment was acquired from the total RNAs of Chinese wild kiwifruit via reverse transcription polymerase chain reaction (RT-PCR), and its DNA sequence was analyzed. Its sequence was determined to share 98.4% homology with the actinidin gene of A. deliciosa. In order to verity the actinidin gene isolated from the Chinese wild kiwifruit in Escherichia coli, the mature gene was amplified via PCR and expressed in E. coli under the control of the T7lac promoter. The actinidin was expressed in E. coli as inclusion bodies, which were solubilized with urea and refolded. The protease activity of the refolded protein was approximately twice as high as that of E. coli BL2l (DE3).

ISOLATION AND CHERACTERIZATION OF ACTINIDIN GENE FROM CHINESE WILD KIWI FRUIT

  • Lee, Nam-Keun;Hahm, Young-Tae
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2000년도 춘계학술발표대회
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    • pp.527-530
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    • 2000
  • 중국산 야생키위로 total RNA를 추출하고 RT-PCR를 실시해서 증폭된 1.2kb fragment를 얻어 pGEM-T Easy에 cloning한 후 염기서열과 아미노산 서열을 비교 분석한 결과 actinidin gene인 것으로 분석되었다.

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