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蜘蛛拖丝蛋白基因亚克隆与全序列测序对其结构与功能了解的意义 被引量:1
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作者 黄建坤 黄智华 李敏 《中国临床康复》 CAS CSCD 2004年第8期1462-1463,T003,共3页
目的:了解蜘蛛拖丝蛋白基因的结构,利用基因工程技术制备人工蛛丝纤维,开发具有独特的机械特性和良好生物相容性的新型蜘蛛丝纤维生物材料。方法:通过结合限制性内切酶消化和超声波机械剪切蜘蛛拖丝蛋白全基因克隆ScosDS-1DNA,制备的DN... 目的:了解蜘蛛拖丝蛋白基因的结构,利用基因工程技术制备人工蛛丝纤维,开发具有独特的机械特性和良好生物相容性的新型蜘蛛丝纤维生物材料。方法:通过结合限制性内切酶消化和超声波机械剪切蜘蛛拖丝蛋白全基因克隆ScosDS-1DNA,制备的DNA片段分别与载体pUC19连接,构建了4个拖丝蛋白基因亚克隆文库。筛选、鉴定亚克隆文库的重组子,选取大小合适的重组子进行测序。结果:筛选的重组子数目超过400个,成功完成的测序反应数近500次,总读长已为ScosDS-1全长的5倍。结论:初步进行了ScosDS-1全序列拼接,以进一步了解拖丝蛋白基因的结构与功能,为合理的设计化合成基因,人工仿制蛛丝纤维奠定了基础。 展开更多
关键词 蜘蛛拖丝蛋白基因 亚克隆 全序列测序 基因工程 生物材料 基因结构
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Genetic Characteristics of 2009 Pandemic H1N1 Influenza A Viruses Isolated from China's Mainland 被引量:4
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作者 Jiu-ru Zhao Yong-dong Li +13 位作者 Li-min Pan Na Zhu Hong-xia Ni Guo-zhang Xu Yong-zhong Jiang Xi-xiang Huo Jun-qiang Xu Han Xia Na Han Shuang Tang Zhong Zhang Zheng Kou Simon Rayner Tian-xian Li 《Virologica Sinica》 CAS CSCD 2011年第6期418-427,共10页
A total of 100 H1N1 flu real-time-PCR positive throat swabs collected from fever patients in Zhejiang, Hubei and Guangdong between June and November 2009, were provided by local CDC laboratories. After MDCK cell cultu... A total of 100 H1N1 flu real-time-PCR positive throat swabs collected from fever patients in Zhejiang, Hubei and Guangdong between June and November 2009, were provided by local CDC laboratories. After MDCK cell culture, 57 Influenza A Pandemic (H1N1) viruses were isolated and submitted for whole genome sequencing. A total of 39 HA sequences, 52 NA sequences, 36 PB2 sequences, 31 PB1 sequences, 40 PA sequences, 48 NP sequences, 51 MP sequences and 36 NS sequences were obtained, including 20 whole genome sequences. Sequence comparison revealed they shared a high degree of homology (96%-99%) with known epidemic strains (A/Califomia/04/2009(H1N1). Phylogenetic analysis showed that although the sequences were highly conserved, they clustered into a small number of groups with only a few distinct strains. Site analysis revealed three substitutions at loop 220 (221-228) of the HA receptor binding site in the 39 HA sequences: A/Hubei/86/2009 PKVRDQEG→PKVRDQEA, A/Zhejiang/08/2009 PKVRDQEG→PKVRDQER, A/Hubei/75/2009 PKVRDQEG→PKVRDQGG, the A/Hubei/75/2009 was isolated from an acute case, while the other two were from patients with mild symptoms. Other key sites such as 119, 274, 292 and 294 amino acids of NA protein,627 of PB2 protein were conserved. Meanwhile, all the M2 protein sequences possessed the Ser32Asn mutation, suggesting that these viruses were resistant to adamantanes. Comparison of these sequences with other H1N1 viruses collected from the NCBI database provides insight into H1N1 transmission and circulation patterns. 展开更多
关键词 H1N1 Phylogenetic analysis Key sites MUTATIONS
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High-throughput sequencing exclusively identified a novel Torque teno virus genotype in serum of a patient with fatal fever 被引量:4
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作者 Zhiqiang Mi Xin Yuan +8 位作者 Guangqian Pei Wei Wang Xiaoping An Zhiyi Zhang Yong Huang Fan Peng Shasha Li Changqing Bai Yigang Tong 《Virologica Sinica》 CAS CSCD 2014年第2期112-118,共7页
Torque teno virus(TTV) has been found to be prevalent world-wide in healthy populations and in patients with various diseases, but its etiological role has not yet been determined. Using high-throughput unbiased seque... Torque teno virus(TTV) has been found to be prevalent world-wide in healthy populations and in patients with various diseases, but its etiological role has not yet been determined. Using high-throughput unbiased sequencing to screen for viruses in the serum of a patient with persistent high fever who died of suspected viral infection and prolonged weakness, we identified the complete genome sequence of a TTV(isolate Hebei-1). The genome of TTV-Hebei-1 is 3649 bp in length, encoding four putative open reading frames, and it has a G+C content of 49%. Genomic comparison and a BLASTN search revealed that the assembled genome of TTV-Hebei-1 represented a novel isolate, with a genome sequence that was highly heterologous to the sequences of other reported TTV strains. A phylogenetic tree constructed using the complete genome sequence showed that TTV-Hebei-1 and an uncharacterized Taiwan Residents strain, TW53A37, constitute a new TTV genotype. The patient was strongly suspected of carrying a viral infection and died eventually without any other possible causes being apparent. No virus other than the novel TTV was identified in his serum sample. Although a direct causal link between the novel TTV genotype infection and the patient's disease could not be confirmed, the findings suggest that surveillance of this novel TTV genotype is necessary and that its role in disease deserves to be explored. 展开更多
关键词 Torque teno virus GENOME persistent high fever high-throughput sequencing
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乙型脑炎病毒减毒活疫苗株(SA14-14-2)神经毒力减弱的分子基础 被引量:7
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作者 李静 俞永新 +2 位作者 安琪 孔燕 杨立宏 《中国病毒病杂志》 CAS 2011年第1期51-57,共7页
目的将乙型脑炎病毒减毒活疫苗株SA14-14-2经乳鼠脑内回传毒力回复后,对回复后病毒测其神经毒力和全长基因序列并与减毒株SA14-14-2和母株SA14进行核苷酸和氨基酸序列比较。方法将回传一代SA14-14-2 M1和SA14-14-2 M3病毒在幼鼠脑内测... 目的将乙型脑炎病毒减毒活疫苗株SA14-14-2经乳鼠脑内回传毒力回复后,对回复后病毒测其神经毒力和全长基因序列并与减毒株SA14-14-2和母株SA14进行核苷酸和氨基酸序列比较。方法将回传一代SA14-14-2 M1和SA14-14-2 M3病毒在幼鼠脑内测神经毒力和全基因组序列。运用RT-PCR方法,分段扩增出覆盖基因组全长的基因片段,PCR产物纯化后直接测序,针对5′和3′端的片段TA克隆后进行测序。经核苷酸序列测定后,利用DNAStar软件,进行序列比对分析。结果实验结果表明SA14-14-2 M1病毒毒力未见增强,SA14-14-2 M3病毒的脑内致病力明显增强,但皮下接种仍无致病性。通过序列比对分析发现,M1和M3病毒均在非结构区的NS3-343、NS4A-57位发生氨基酸回复突变,但只有M3病毒在E区毒力相关的E107、E138位点发生回复突变。结论在SA14-14-2减毒株病毒结构基因中E蛋白区E107、E138位氨基酸是与毒力密切相关的重要位点,应该是神经毒力减弱的重要分子基础,非结构区的个别位点对神经毒力减弱也可能起一定的协同作用。 展开更多
关键词 乙型脑炎病毒 乳鼠脑内回传 神经毒力 回复突变 基因全序列测序
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Genomes and evolutionary genomics of animals
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作者 Luting SONG Wen WANG 《Current Zoology》 SCIE CAS CSCD 2013年第1期87-98,共12页
Alongside recent advances and booming applications of DNA sequencing technologies, a great number of complete genome sequences for animal species are available to researchers. Hundreds of animals have been involved in... Alongside recent advances and booming applications of DNA sequencing technologies, a great number of complete genome sequences for animal species are available to researchers. Hundreds of animals have been involved in whole genome se- quencing, and at least 87 non-human animal species' complete or draft genome sequences have been published since 1998. Based on these technological advances and the subsequent accumulation of large quantity of genomic data, evolutionary genomics has become one of the most rapidly advancing disciplines in biology. Scientists now can perform a number of comparative and evolu- tionary genomic studies for animals, to identify conserved genes or other functional elements among species, genomic elements that confer animals their own specific characteristics and new phenotypes for adaptation. This review deals with the current ge- nomic and evolutionary research on non-human animals, and displays a comprehensive landscape of genomes and the evolution- ary genomics of non-human animals. It is very helpful to a better understanding of the biology and evolution of the myriad forms within the animal kingdom [Current Zoology 59 (1): 87-98, 2013]. 展开更多
关键词 ANIMALS GENOME Evolutionary genomics
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