检索结果-南通市图书馆

Arginine kinase in Toxocara canis:Exon-intron organization,functional analysis of site-directed mutants and evaluation of putative enzyme inhibitors

引用

Asian Pacific Journal of Tropical Medicine 2016年第10期9卷 973-979页

作者： Susiji Wickramasinghe lalani yatawara Mitsuru Nagataki Takeshi Agatsuma Department of Parasitology Faculty of MedicineUniversity of Peradeniya Department of Medical Laboratory sciences Faculty of Allied Health SciencesUniversity of Peradeniya Department of Environmental Health Sciences Kochi Medical School

Objective: To determine exon/intron organization of the Toxocara canis(T. canis) AK(TCAK) and to test green and black tea and several other chemicals against the activity of recombinant TCAK in the guanidino-specific region by site-directed mutants. Methods: Amplification of genomic DNA fragments containing introns was carried out by PCRs. The open-reading frame(1 200 bp) of TCAK(wild type) was cloned into the BamH 1/SalI site of pM AL-c2X. The maltose-binding protein-TCAK fusion protein was expressed in Escherichia coli TB1 cells. The purity of the expressed enzyme was verified by SDS-PAGE. Mutations were introduced into the guanidino-specific region and other areas of pM AL/TCAK by PCR. Enzyme activity was measured with an NADH-linked assay at 25℃ for the forward reaction(phosphagen synthesis). Results: Arginine kinase in T. canis has a seven-exon/six-intron gene structure. The lengths of the introns ranged from 542 bp to 2 500 bp. All introns begin with gt and end with ag. Furthermore, we measured the enzyme activity of site-directed mutants of the recombinant TCAK. The K_m value of the mutant(Alanine to Serine) decreased indicating a higher affinity for substrate arginine than the wild-type. The K_m value of the mutant(Serine to Glycine) increased to 0.19 mM. The Km value(0.19 mM) of the double mutant(Alanine-Serine to Serine-Glycine) was slightly greater than in the wild-type(0.12 mM). In addition, several other chemicals were tested; including plant extract Azadiracta indica(A. indica), an aminoglycoside antibiotic(aminosidine), a citrus flavonoid glycoside(rutin) and a commercially available catechin mixture against TCAK. Green and black tea(1:10 dilution) produced 15% and 25% inhibition of TCAK, respectively. The extract of A. indica produced 5% inhibition of TCAK. Moreover, green and black tea produced a non-competitive type of inhibition and A. indica produced a mixed-type of inhibition on TCAK. Conclusions: Arginine kinase in T. canis has a seven-exon/six

关键词： Toxocara canis Arginine kinase Gene structure Site directed mutagenesis Inhibition kinetics

来源：

维普期刊数据库评论

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

Organochlorine pesticide and heavy metal residues in some edible biota collected from Quanzhou Bay and Xinghua Bay,Southeast China

引用

Journal of Environmental Sciences 2010年第2期22卷 314-320页

作者： Mangala yatawara Odhiambo Joshua Owago Julia E Burnet Key Laboratory of Biogeology and Environmental Geology of Ministry of Education

In order to assess the contamination of edible marine organisms in Quanzhou and Xinghua Bays, Fujian Province, China, concentrations of organochlorine pesticides （hexachlorocyclohexane isomers or HCHs and dichlorodiphenyltrichloroethane isomers/derivatives or DDTs） and heavy metals （Cu, Pb, Zn, As, Cr, Cd, Ni and Hg） in 13 species collected from different sites in the hays were analyzed by GC-ECD and ICP-MS. The concentration of the sum of DDTs exceeded that the sum of HCHs in the samples. Most of the organisms showed higher levels of DDTs than the first level criterion （10 ng/g） for marine biological quality in China （GB 18421-2001） but conformed to the first level criterion （20 ng/g） for HCHs. The estimated mean daily uptake of pesticides was below the level for minimal risk to the consumers. The levels of Cd, As and Pb in most organisms from both bays were markedly elevated and above the recommended legal limits for human consumption. Our results showed the need for routine monitoring of marine species contamination to ensure public and environmental health.

关键词： organochlorine pesticides residue levels technical HCH heavy metals Fujian Province

来源：

维普期刊数据库

同方期刊数据库评论

在线全文

学校读者我要写书评

暂无评论

欢迎您,

建议与咨询留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

时间限定

文献类型

馆藏选择

核心期刊

语言

文献类型

帮助

文字说明：

检索规则说明：

检索范例：

分类表

所选分类

限定检索结果

文献类型

馆藏范围

日期分布

学科分类号

主题

机构

作者

语言

在线全文

在线全文

请选择保存的检索档案：

请选择收藏分类：

通借通还

欢迎您,

建议与咨询 留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

时间限定

文献类型

馆藏选择

核心期刊

语言

文献类型

帮助

文字说明：

检索规则说明：

检索范例：

分类表

所选分类

限定检索结果

文献类型

馆藏范围

日期分布

学科分类号

主题

机构

作者

语言

在线全文

在线全文

请选择保存的检索档案： 新增检索档案 确定 取消

请选择收藏分类： 新增自定义分类 确定 取消

通借通还

建议与咨询留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

请选择保存的检索档案：

请选择收藏分类：