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1.
Tanapan Prommool Pongpawan Sethanant Narodom Phaenthaisong Nattaya Tangthawornchaikul Adisak Songjaeng Panisadee Avirutnan Dumrong Mairiang Prasit Luangaram Chatchawan Srisawat Watchara Kasinrerk Sirijitt Vasanawathana Kanokwan Sriruksa Wannee Limpitikul Prida Malasit Chunya Puttikhunt 《PLoS neglected tropical diseases》2021,15(2)
2.
Moonsom S Chaisri U Kasinrerk W Angsuthanasombat C 《Journal of biochemistry and molecular biology》2007,40(5):783-790
Receptor binding plays an important role in determining host specificity of the Bacillus thuringiensis Cry delta-endotoxins. Mutations in domains II and III have suggested the participation of certain residues in receptor recognition and insect specificity. In the present study, we expressed the cloned domain II-III fragment of Cry4Ba and examined its binding characteristics to mosquito-larval midgut proteins. The 43-kDa Cry4Ba-domain II-III protein over-expressed in Escherichia coli as inclusion bodies was only soluble when carbonate buffer, pH 10.0 was supplemented with 4 M urea. After renaturation via stepwise dialysis and subsequent purification, the refolded domain II-III protein, which specifically reacts with anti Cry4Ba-domain III monoclonal antibody, predominantly exists as a beta-sheet structure determined by circular dichroism spectroscopy. In vitro binding analysis to both histological midgut tissue sections and brush border membrane proteins prepared from susceptible Aedes aegypti mosquito-larvae revealed that the isolated Cry4Ba-domain II-III protein showed binding functionality comparable to the 65-kDa full-length active toxin. Altogether, the data present the 43-kDa Cry4Ba fragment comprising domains II and III that was produced in isolation was able to retain its receptor-binding characteristics to the target larval midgut proteins. 相似文献
3.
Ratchada Cressey Saranya Pimpa Busyamas Chewaskulyong Nirush Lertprasertsuke Somchareon Saeteng Chatchai Tayapiwatana Watchara Kasinrerk 《BMC biotechnology》2008,8(1):16
Background
The recognition that human tumors stimulate the production of autoantibodies has initiated the use of this immune response as serological markers for the early diagnosis and management of cancer. The enzyme-linked immunosorbent assay (ELISA) is the most common method used in detecting autoantibodies, which involves coating the microtiter plate with the tumor associated antigen (TAA) of interest and allowing serum antibodies to bind. The patient's sample is directly in contact with the coating antigen so the protein used for coating must be pure to avoid non-specific binding. In this study, a simplified method to selectively and specifically immobilize TAAs onto microtiter plates in order to detect circulating autoantibodies in cancer patients without prior purification process was described. Wild type full-length p53 protein was produced in fusion with biotin carboxyl carrier peptide (BCCP) or hexahistidine [(His)6] using pAK400 and pET15b(+) vectors, respectively. The recombinant p53 fusion protein produced was then subjected to react with either a commercial p53 monoclonal antibody (mAb) or sera from lung cancer patients and healthy volunteers in an enzyme-linked immunosorbent assay (ELISA) format. 相似文献4.
Porntip Paopang Watchara Kasinrerk Chatchai Tayapiwatana Phisit Seesuriyachan 《Preparative biochemistry & biotechnology》2016,46(3):305-312
The single-chain fragment variable (scFv) was used to produce a completely functional antigen-binding fragment in bacterial systems. The advancements in antibody engineering have simplified the method of producing Fv fragments and made it more efficient and generally relevant. In a previous study, the scFv anti HIV-1 P17 protein was produced by a batch production system, optimized by the sequential simplex optimization method. This study continued that work in order to enhance secreted scFv production by fed-batch cultivation, which supported high volumetric productivity and provided a large amount of scFvs for diagnostic and therapeutic research. The developments in cell culture media and process parameter settings were required to realize the maximum production of cells. This study investigated the combined optimization methods, Plackett–Burman design (PBD) and sequential simplex optimization, with the aim of optimize feed medium. Fed-batch cultivation with an optimal feeding rate was determined. The result demonstrated that a 20-mL/hr feeding rate of the optimized medium can increase cell growth, total protein production, and scFv anti-p17 activity by 4.43, 1.48, and 6.5 times more than batch cultivation, respectively. The combined optimization method demonstrated novel power tools for the optimization strategy of multiparameter experiments. 相似文献
5.
Saichit Khummuang Waraporn Phanphrom Witida Laopajon Watchara Kasinrerk Ponlatham Chaiyarit Supansa Pata 《Biological procedures online》2017,19(1):14
Background
Human trefoil factor (TFF) peptides consist of three members: TFF1, TFF2 and TFF3. TFF3 is the most abundant TFF peptide in saliva. TFF3 homodimer was suggested to be involved in apoptosis inhibition and malignancy. Determination of TFF3 homodimer expression profiles in saliva may lead to new information about oral biology and diseases. The objective of this study was to generate monoclonal antibodies (mAbs) against TFF3 and apply the produced mAbs for the establishment of ELISA for quantification of dimeric TFF3 in saliva.Results
With our modified hybridoma technique, three hybridoma clones producing anti-TFF3 mAbs having IgG isotype were generated. The mAbs were specific for TFF3 with no cross-reactivity to other TFFs. Using the generated mAbs, a modified-sandwich ELISA with high sensitivity for the quantification of dimeric TFF3 in saliva was developed. Using this ELISA, the amount of dimeric TFF3 in saliva could be measured.Conclusions
A modified-sandwich ELISA for the quantification of TFF3 dimeric form was established. The established ELISA will be a valuable tool for facilitating the investigation of the physiological roles and the diagnostic values of TFF3 in oral diseases. The concept of this modified-sandwich ELISA may be applied for the determination of other homodimeric peptides of interest.6.
Suriyan Cha-um Chaturong Chanseetis Watchara Chintakovid Aussanee Pichakum Kanyaratt Supaibulwatana 《Plant Cell, Tissue and Organ Culture》2011,106(3):435-444
In this study, a rooting protocol was developed for macadamia plantlets with healthy roots and enhanced growth performance,
along with enhanced photosynthetic capability. In vitro-grown shoots rooted in vented vessels containing vermiculite as the
supporting material exhibited 100% frequency of root induction, whereas when shoots were grown in non-vented vessels containing
a solidified Murashige and Skoog (MS) medium, the frequency of root induction was less than 30%. The formation of root with
callus, hyperhydricity, and leaf necrosis was observed in this photomixotrophic closed system. The modification of the vented
photoautotrophic system with different concentrations of CO2 and sucrose were investigated using vermiculite as the supporter. The number of roots, root length, root surface area, fresh
weight, and dry weight were significantly higher in plantlets grown in CO2-enriched (1,000 μmol CO2 mol−1) photoautotrophic conditions. The water content in both root and shoot tissues of plantlets cultured under photoautotrophic
conditions was maximized. In addition, shoot and leaf performances were enhanced in plantlets cultured under CO2-enriched photoautotrophic conditions. The supplementation of sucrose (29–88 mM) to culture media in both ambient and elevated
CO2 conditions affected a reduction in the shoot and root performance of in vitro plantlets. Chlorophyll a, chlorophyll b, and
total carotenoids in the leaf tissues of plantlets acclimatized in CO2-enriched photoautotrophic conditions were enriched, leading to increasing photosynthetic abilities, including chlorophyll
fluorescence and net photosynthetic rate. From this investigation, a root induction protocol was established and the production
of healthy macadamia plantlets was successfully implemented using CO2-enriched photoautotrophic conditions. 相似文献
7.
Yufei Dai Rachawadee Chantra Kongkiat Kittiwattanawong Liyuan Zhao Watchara Sakornwimon Reyilamu Aierken Fuxing Wu Xianyan Wang 《Genetics and molecular biology》2021,44(2)
The Irrawaddy dolphin (Orcaella brevirostris) is an endangered, small cetacean species which is widely distributed in rivers, estuaries, and coastal waters throughout the tropical and subtropical Indo-Pacific. Despite the extensive distribution of this species, little is known of individual movements or genetic exchange among regions in Thailand. Here, we evaluate the genetic diversity and genetic structure of O. brevirostris in the eastern, northern and western Gulf of Thailand, and Andaman Sea. Although phylogenetic relationships and network analysis based on 15 haplotypes obtained from 32 individuals reveal no obvious divergence, significant genetic differentiation in mitochondrial DNA (overall FST = 0.226, P < 0.001; ΦST = 0.252, P < 0.001) is apparent among regions. Of 18 tested microsatellite loci, 10 are polymorphic and successfully characterized in 28 individuals, revealing significant genetic differentiation (overall FST = 0.077, P < 0.05) among the four sampling sites. Structure analysis reveals two inferred genetic clusters. Additionally, Mantel analysis demonstrates individual-by-individual genetic distances and geographic distances follow an isolation-by-distance model. We speculate that the significant genetic structure of O. brevirostris in Thailand is associated with a combination of geographical distribution patterns, environmental and anthropogenic factors, and local adaptations. 相似文献
8.
Characterization of thermotolerant Acetobacter pasteurianus strains and their quinoprotein alcohol dehydrogenases 总被引:2,自引:0,他引:2
Watchara Kanchanarach Gunjana Theeragool Toshiharu Yakushi Hirohide Toyama Osao Adachi Kazunobu Matsushita 《Applied microbiology and biotechnology》2010,85(3):741-751
We isolated several thermotolerant Acetobacter species of which MSU10 strain, identified as Acetobacter pasteurianus, could grow well on agar plates at 41°C, tolerate to 1.5% acetic acid or 4% ethanol at 39°C, similarly seen with A. pasteurianus SKU1108 previously isolated. The MSU10 strain showed higher acetic acid productivity in a medium containing 6% ethanol at 37°C than SKU1108 while SKU1108 strain could accumulate more acetic acid in a medium supplemented with 4–5% ethanol at the same temperature. The fermentation ability at 37°C of these thermotolerant strains was superior to that of mesophilic A. pasteurianus IFO3191 strain having weak growth and very delayed acetic acid production at 37°C even at 4% ethanol. Alcohol dehydrogenases (ADHs) were purified from MSU10, SKU1108, and IFO3191 strains, and their properties were compared related to the thermotolerance. ADH of the thermotolerant strains had a little higher optimal temperature and heat stability than that of mesophilic IFO3191. More critically, ADHs from MSU10 and SKU1108 strains exhibited a higher resistance to ethanol and acetic acid than IFO3191 enzyme at elevated temperature. Furthermore, in this study, the ADH genes were cloned, and the amino acid sequences of ADH subunit I, subunit II, and subunit III were compared. The difference in the amino acid residues could be seen, seemingly related to the thermotolerance, between MSU10 or SKU1108 ADH and IFO 3191 ADH. 相似文献
9.
Laenoi W Rangkasenee N Uddin MJ Cinar MU Phatsara C Tesfaye D Scholz AM Tholen E Looft C Mielenz M Sauerwein H Wimmers K Schellander K 《Molecular biology reports》2012,39(4):3893-3901
The present study was aimed to determine the association between metalloproteinase 3 (MMP3), transforming growth factor beta 1 (TGF??1) and collagen type X alpha I (COL10A1) gene polymorphisms with traits related to leg weakness in pigs. Three hundred Duroc?×?Pietrain cross breds (DuPi) and 299 pigs of a commercial population (CP) were used for the experiment. DuPi animals were examined for 10 different traits describing leg and feet structure, osteochondrosis (OC) scores and bone density status. Data of OC score at condylus medialis humeri, condylus medialis femoris and distal epiphysis ulna regions of CP were used for association analysis. Significant association (P?<?0.05) was found for MMP3 SNP (g.158 C>T) with OC at head of femur and bone mineral density in the DuPi population. Association (P?<?0.05) was found between SNP of TGF??1 (g.180 G>A) with rear leg score and the principle component denoting both OC and feet and leg scores in the DuPi population. No association was found between COL10A1 (g.72 C>T) and leg weakness related traits. The associations of SNPs with OC traits could not be confirmed in the commercial population. Expression analysis of the three candidate genes was performed to compare between healthy and OC. TGF??1 was found to be highly expressed (P?<?0.05) in the OC compared to healthy cartilages, but no significant different expressions were observed for MMP3 and COL10A1 genes. The present finding suggested that TGF??1 and MMP3 genes variants have an effect on some of the leg weakness related traits. 相似文献
10.
Keelapang P Sriburi R Supasa S Panyadee N Songjaeng A Jairungsri A Puttikhunt C Kasinrerk W Malasit P Sittisombut N 《Journal of virology》2004,78(5):2367-2381
During the export of flavivirus particles through the secretory pathway, a viral envelope glycoprotein, prM, is cleaved by the proprotein convertase furin; this cleavage is required for the subsequent rearrangement of receptor-binding E glycoprotein and for virus infectivity. Similar to many furin substrates, prM in vector-borne flaviviruses contains basic residues at positions P1, P2, and P4 proximal to the cleavage site; in addition, a number of charged residues are found at position P3 and between positions P5 and P13 that are conserved for each flavivirus antigenic complex. The influence of additional charged residues on pr-M cleavage and virus replication was investigated by replacing the 13-amino-acid, cleavage-proximal region of a dengue virus (strain 16681) with those of tick-borne encephalitis virus (TBEV), yellow fever virus (YFV), and Japanese encephalitis virus (JEV) and by comparing the resultant chimeric viruses generated from RNA-transfected mosquito cells. Among the three chimeric viruses, cleavage of prM was enhanced to a larger extent in JEVpr/16681 than in YFVpr/16681 but was slightly reduced in TBEVpr/16681. Unexpectedly, JEVpr/16681 exhibited decreased focus size, reduced peak titer, and depressed replication in C6/36, PS, and Vero cell lines. The reduction of JEVpr/16681 multiplication correlated with delayed export of infectious virions out of infected cells but not with changes in specific infectivity. Binding of JEVpr/16681 to immobilized heparin and the heparin-inhibitable infection of cells were not altered. Thus, diverse pr-M junction-proximal sequences of flaviviruses differentially influence pr-M cleavage when tested in a dengue virus prM background. More importantly, greatly enhanced prM cleavability adversely affects dengue virus export while exerting a minimal effect on infectivity. Because extensive changes of charged residues at the pr-M junction, as in JEVpr/16681, were not observed among a large number of dengue virus isolates, these results provide a possible mechanism by which the sequence conservation of the pr-M junction of dengue virus is maintained in nature. 相似文献