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Cloning of Banana Bunchy Top Virus Chinese Zhangzhou Isolate DNA 4 and the Promoter Activity of Its Non_coding Region
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作者 孙德俊 魏红艳 +1 位作者 蔡文启 田颖川 《Acta Botanica Sinica》 CSCD 2002年第8期941-945,共5页
Banana bunchy top virus Chinese Zhangzhou isolate (BBTV-ZZ) DNA 4 was amplified by PCR and cloned. Sequence analysis showed that BBTV-ZZ DNA 4 is 1 039 nucleotides (nts) in length and this virus could be one member of... Banana bunchy top virus Chinese Zhangzhou isolate (BBTV-ZZ) DNA 4 was amplified by PCR and cloned. Sequence analysis showed that BBTV-ZZ DNA 4 is 1 039 nucleotides (nts) in length and this virus could be one member of BBTV Asian group. Transcriptional initiation site A, which is at the 269 nucleotide, was preliminarily determined by using 5' RACE method. BBTV-ZZ DNA 4 non-coding region was sub-cloned by PCR and inserted into upstream of gfp : : gus plant expression vector pCAMBIA 1304 to construct recombinant plasmid pTA2. Agrobacterium tumefaciens harboring pTA2 was injected into leaves of the tobacco (Nicotiana tabacum L. cv. Xanthi NC) via Agrobacterium-infiltration procedure. Transient expressions of GUS and GFP were determined in injected leaves 3 - 5 d later. GUS activities of pTA2, pCAMBIA 1304 injected and non-injected tobacco leaves respectively were 1.007 0 pmol MU(.)mug(-1.)min(-1), 2.069 0 pmol MU(.)mug(-1.)min(-1) and 0.021 4 pmol MU(.)mug(-1.)min(-1). Indirect ELISA for GFP in 1 mg total protein from pTA2, pCAMBIA 1304 injected and non-injected leaves showed an A(490 nm) value of 89.577, 100.440 and 3.287, respectively. These results showed that the non-coding region of BBTV-ZZ DNA 4 has a promoter activity not only in the virus replication in monocot, but also in driving the expression of a foreign gene in dicot plants. 展开更多
关键词 banana bunchy top virus Chinese Zhangzhou isolate DNA 4 5 ' RACE PROMOTER GFP GUS
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“克隆”在医疗技术中的应用 被引量:3
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作者 胡作怀 王晓华 《中外医用放射技术》 2001年第6期3-3,共1页
关键词 医疗技术 “硬盘克隆软件” 医疗设备 “克隆分区” X线设备
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Difference in DNA sequences in SSU rDNA variable regions among pathogens isola ted from different epidemic foci of visceral leishmaniasis in China 被引量:1
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作者 胡孝素 卜玲毅 +3 位作者 马莹 王雅静 敬保迁 易桃林 《Chinese Medical Journal》 SCIE CAS CSCD 2002年第10期1457-1459,共3页
OBJECTIVE: To confirm the existence of point mutations in the SSU rDNA variable regions of 5 Leishmania donovani (L.d.) isolates from different epidemic foci in China. METHODS: Specific SSU rDNA fragments from nuclear... OBJECTIVE: To confirm the existence of point mutations in the SSU rDNA variable regions of 5 Leishmania donovani (L.d.) isolates from different epidemic foci in China. METHODS: Specific SSU rDNA fragments from nuclear DNA of 7 Leishmania species/isolates were amplified by PCR and then cloned into pGEM(R)-T Easy Vectors. After that, the specific fragments were sequenced by an automated DNA sequencer. RESULTS: Sequence analysis showed that the amplified DNA fragments of 7 Leishmania species/isolates were all 392 bp in length. All 5 point mutations were located in two unique sequence blocks (UQ-I and UQ-II), and no insertions or deletions were found. The identities of comparison of Leishmania in GeneBank were more than 98%. CONCLUSION: Five point mutations exist in the SSU rDNA variable region of 5 L.d. isolates from different epidemic foci of visceral leishmaniasis (VL) in China. Sequence differences of the SSU rDNA variable region exist among L.d. isolates from different foci. 展开更多
关键词 Point Mutation Animals DNA Protozoan DNA Ribosomal Humans Leishmania donovani Leishmaniasis Visceral Polymerase Chain Reaction Research Support Non-U.S. Gov't
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Difference in DNA sequences in SSU rDNA variable regions among pathogens isolated from different epidemic foci of visceral leishmaniasis in China
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作者 胡孝素 卜玲毅 +3 位作者 马莹 王雅静 敬保迁 易桃林 《Chinese Medical Journal》 SCIE CAS CSCD 2002年第10期17-19,143,共4页
To confirm the existence of point mutations in the SSU rDNA variable regions of 5 Leishmania donovani (L d ) isolates from different epidemic foci in China Methods Specific SSU rDNA fragments from nuclear DNA of 7... To confirm the existence of point mutations in the SSU rDNA variable regions of 5 Leishmania donovani (L d ) isolates from different epidemic foci in China Methods Specific SSU rDNA fragments from nuclear DNA of 7 Leishmania species/isolates were amplified by PCR and then cloned into pGEM R T Easy Vectors After that, the specific fragments were sequenced by an automated DNA sequencer Results Sequence analysis showed that the amplified DNA fragments of 7 Leishmania species/isolates were all 392 bp in length All 5 point mutations were located in two unique sequence blocks (UQ Ⅰ and UQ Ⅱ), and no insertions or deletions were found The identities of comparison of Leishmania in GeneBank were more than 98% Conclusion Five point mutations exist in the SSU rDNA variable region of 5 L d isolates from different epidemic foci of visceral leishmaniasis (VL) in China Sequence differences of the SSU rDNA variable region exist among L d isolates from different foci 展开更多
关键词 visceral leishmaniasis · Leishmania · point mutation · small subunit ribosomal DNA · polymerase chain reaction · cloning · sequence analysis · epidemic foci
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