A Ferritin from Dendrorhynchus zhejiangensis with Heavy Metals Detoxification Activity
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Ferritin, an iron homeostasis protein, has important functions in transition and storage of toxic metal ions. In this study, the full-length cDNA of ferritin was isolated from Dendrorhynchus zhejiangensis by cDNA library and RACE approaches. The higher similarity and conserved motifs for ferritin were also identified in worm counterparts, indicating that it belonged to a new member of ferritin family. The temporal expression of worm ferritin in haemocytes was analyzed by RT-PCR, and revealed the ferritin could be induced by Cd2+, Pb2+ and Fe2+. The heavy metal binding activity of recombinant ferritin was further elucidated by atomic force microscopy (AFM). It was observed that the ferritin protein could form a chain of beads with different size against three metals exposure, and the largest one with 35∼40 nm in height was identified in the Cd2+ challenge group. Our results indicated that worm ferritin was a promising candidate for heavy metals detoxification.Keywords:
Detoxification
Objective To study the characteristics of expression of self-cloned 1.5 kb hLYZ cDNA.Methods Subclone the 1.5 kb hLYZ cDNA into an eukaryotic expression vector pcDNA3 and mammary gland expression vector p205C3.Transfect COS-1 cells with recombinant plasmid pcDNA3LYZ and detect the expression of hLYZ cDNA by IFA.Inject the mice during lactating period with recombinant plasmid p205C3LYZ and detect the expression of hLYZ cDNA in vivo of mice by micrococcal lysis assay and dot blot.Results hLYZ cDNA was correctly expressed in the transfected COS-1 cells.The mean expression level of hLYZ in milk of mice was 87 μg/ml.The expression of target protein was tissue-specific.Conclusion The self-cloned hLYZ cDNA may be used for the preparation and development of recombinant hLYZ.The recombinant plasmid p205C3LYZ may be used for the preparation of animal mammary gland bioreactors.
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In this study, 20 subjects who had home detoxification with supervision and support from the Western Australian Alcohol and Drug Authority Community Nursing Service were matched with 20 subjects who had inpatient detoxification in the Authority's detoxification facility. Subjects were interviewed between nine and 22 months (mean 15.5 months) after detoxification to compare client outcomes and the costs of home and inpatient detoxification. The results indicate that, for suitable clients, home detoxification was at least as beneficial as inpatient detoxification and that it was achieved at a much lower cost than inpatient care.
Detoxification
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Detoxication
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Based on the present status of chromium containing slag′s detoxification,the detoxification mechanisms,specifications and the feasibility of the methods,including chemical detoxification,detoxification by microwave radiating and biological detoxification,are introduced and compared.Finally the prospect of the technology on detoxification of the chromium containing slag is also proposed.
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Expression of human interleukin 2 (IL-2) at high levels has been achieved in human cells by cotransfection and subsequent coamplification of the transfected sequences. IL-2 production by these cells reached 2700 U/ml of culture medium. This productivity was higher than the 100U/ml attained by concanavalin A (ConA) stimulated human leukaemic Jurkat, T cells. A plasmid, pSDI, containing human IL-2 cDNA and mouse dihydrofolate reductase (DHFR) cDNA was constructed, and cotransfected with plasmid pSV2neo at a 10:1 molar ratio, into Hela (human epitheloid carcinoma) cells by the DNA-calcium phosphate coprecipitation technique. Transformants producing IL-2 were easily obtained among antibiotic G418 resistant colonies. Cells resistant to increased levels of methotrexate (MTX) secreted a large amount of IL-2 as a result of coamplification of DHFR cDNA and IL-2 cDNA. Expression of IL-2 cDNA was stimulated by removing a G-C stretch which was present between Simian Virus 40 (SV40) early promoter and IL-2 cDNA. Plasmid pSDI was rescued from the transformed Hela cells and shown to be present in the intact form among the amplified cellular DNA.
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genomic DNA
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A cDNA clone for the 90kDa heat-shock protein, which we have recently identified as a component of steroid hormone receptors in their heteromeric 8S form, was isolated by direct immunological screening of a chicken smooth muscle cDNA expression library, prepared in the expression plasmids pUC8 and pUC9. Using polyclonal and monoclonal antibodies against the 90kDa protein a colony was identified that reacted with both antibodies. Plasmid 9.11 (p9.11, ˜ 1100 base pair insert) was found to hybrid-select mRNA for the 90kDa heat-shock protein. Northern blot analysis revealed that RNA isolated from various chicken tissues contain a single transcript of ˜ 3 Kb hybridizing to a [32P]labelled cDNA probe made from p9.11. Heat-shock treatment of chick embryonic fibroblasts resulted in increased steady-state levels of a 3 Kb transcript in both poly A+ and poly A− RNA fractions. Southern blot analysis of chicken genomic DNA indicated that the cDNA hybridizes to a single copy sequence. Sequence data show that the p9.11 cDNA displays a high degree of homology with the 5′ portion of yeast heat shock protein 90 cDNA.
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We have isolated and analyzed cDNA (designated P ‐450 HP cDNA) clones from a human placenta cDNA library, using the cDNA for rabbit pulmonary cytochrome P ‐450 P‐2 , a prostaglandin ω‐hydroxylase, as a hybridization probe. The cDNA obtained encoded a polypeptide comprising 511 amino acids with a calculated molecular mass of 58987 Da, and the amino acid sequence similarity with P ‐450 P‐2 and rat liver laurate ω‐hydroxylase ( P ‐450 LA ω) was only about 50%. RNA blot analysis showed that the mRNA hybridizable with the human P ‐450 HP cDNA was inducibly expressed 3 – 5‐fold in rabbit small intestine and lung by gestation, but the expression remained constant in rabbit liver and kidney. This mode of expression was quite different from that of P ‐450 P‐2 and P ‐450 LA ω. Interestingly, the mRNA hybridized with the cDNA of P ‐450 HP was found to be expressed in all the human tumor tissues so far examined, in sharp contrast with the facts that almost all the other species of P ‐450 s are known to disappear in the tumor tissues. Taken together, the deduced hemoprotein termed P ‐450 HP dose not seem to be the human counterpart of rabbit P ‐450 P‐2 or rat P ‐450 LA ω, and is presumably a new member of the P ‐450 family including P ‐450 P‐2 and P ‐450 LA ω. Furthermore, the corresponding genomic DNA was also cloned and analyzed. The gene of P ‐450 HP spanned 18.8 kb and was separated into 11 exons by 10 introns whose locations were completely different from those of P ‐450 genes so far determined.
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Abstract One intermediate treatment goal, and a preliminary phase of those treatments that are aimed at abstinence involves withdrawal from drugs or ‘detoxification’.1
Detoxification
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Detoxification
Opiate
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