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Construction of neuron specific vector of human antisense noggin gene expression

Neural Regeneration Research 2010年第15期5卷 1171-1174页

作者： Shengnian Zhou Chengshan Li Xiansen Wei Liqing Liu Zhengda Zhang Department of Neurology Qilu Hospital of Shandong University Jinan 250012 Shandong Province China Department of Cadre Medicine China-Japan Friendship Hospital Beijing 100029 China Department of Basic Medicine Shandong University of Traditional Chinese Medicine Jinan 250355 Shandong Province China

Thenoggin gene is present in the central nervous system in embryonic and postnatal mammals, and plays an important role in maintaining nervous system development and physiological function A 0.76-kb sequence of human noggin gene was cloned by polymerase chain reaction with the digestion site of Hind Ill and Xba I on the 5＇ end. The cloned fragment was reversely inserted into pCS2＋[Tal]-GFP plasmid, an neural cell-specific antisense eukaryotic expression vector. The plasmid expresses antisense for human noggin specifically in neurons, which may facilitate understanding of the physiological function of noggin.

关键词： noggin alphal-tubulin expression vector antisense nucleic acids nerve cells neural regeneration

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Construction of Prokaryotic Expressing Vector of antisense nucleic Acid of lasR and Its Effect on the Virulence of Pseudomonas Aeruginosus

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2007年第4期27卷 349-353页

作者：张玲周俊立李静铭廖芳 Department of Pathogenic Biology Tongji Medical CollegeHuazhong University of Science and Technology

To construct a pUCP18/lasR^antisense plasmid carrying the reversed gene and analyze its effect on the virulence of Pseudomonas aeruginosus, LasR gene was amplified from the genome of Pseudomonas aeruginosus by PCR and reversely recombined with plasmid pUCP18. The recombinant pUCP18/lasR^antisense was verified by enzyme digestion, PCR and sequencing. The biological effects of pUCP18/lasR^antisense were examined by using RT-PCR, NAD method and the assay of pyocyanin. Our results showed that the expected full length lasR fragment （721 bp） was extended from Pseudomonas aeruginosus gene with PCR. And it is consistent with LasR gene of Pseudomonas aeruginosa in GenBank （No. NC_002516）. The recombinant plasmid was successfully constructed and transferred into Pseudomonas aeruginosus. The antisense nucleic acid of LasR gene could reduce the virulence of Pseudomonas aeruginosus and might serve as a new target site for treatment purpose.

关键词： LasR gene antisense nucleic acids bacterial transformation virulence factor

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