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中国生物工程杂志

China Biotechnology
China Biotechnology
China Biotechnology  2011, Vol. 31 Issue (06): 99-105    DOI:
    
The Design and in vitro Transcription of pRNA-Ribozymes
ZHOU Ying1,2, FANG Jing2, MAO Jian-ping2, ZHU Yan1
1. Basic Medicine College, Anhui Medical University, Hefei 230032, China;
2. Beijing Institute of Radiation Medicine, Beijing 100850, China
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Abstract  

Objectives: pRNA, the packaging-RNA molecule of Bacillus subtilis is a new nanometer-molecular carrier. The recombination of pRNA and the hammerhead Ribozyme can be used to construct stable, self-deliverable pRNA-Ribozymes for entering cells and actively binding and cutting RNA. However, it is technically difficult to synthesize RNA molecules longer than 100 nt by chemistry. Therefore, 170 nt pRNA-Ribozyme was prepared by genetic recombination and in vitro transcription. Methods: The DNA of pRNA and Ribozyme were amplified by PCR before recombination according to their molecule sequences. The pRNA-Ribozyme was reconstructed in two forms: the chimeric ribozyme linked to pRNA vector as a cis-connected open structure, and the chimeric ribozyme harbored in pRNA vector as a closing inner structure. The pRNA-Ribozymes were obtained by in vitro transcription. Results: The pRNA-Ribozyme of cis-connected open structure and the embedded inner structure were prepared by in vitro transcription. Both molecules could form the polymer structure in vitro through correct folding. The pRNA-Ribozyme had the activity for binding and cutting its target mRNAs, which provides the foundation for further application of pRNA-Ribozyme in intracellular.

Key words： pRNA      Ribozyme     
Received: 25 February 2011      Published: 28 June 2011
ZTFLH:  Q819  
Corresponding Authors: Peter JMao     E-mail: maojp@nic.bmi.ac.cn; zhuyan@ah.edu.cn
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ZHOU Ying
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ZHOU Ying, FANG Jing, MAO Jian-ping, ZHU Yan. The Design and in vitro Transcription of pRNA-Ribozymes. China Biotechnology, 2011, 31(06): 99-105.

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https://manu60.magtech.com.cn/biotech/     OR     https://manu60.magtech.com.cn/biotech/Y2011/V31/I06/99

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