| high-level expression of a soluble snake venom enzyme, gloshedobin, in e. coli in the presence of metal ions. | the mature gene of gloshedobin, a snake venom thrombin-like enzyme from the snake, gloydius shedaoensis, was cloned and expressed in strain e. coli bl21 (de3). having been induced by iptg, the recombinant gloshedobin was in both soluble and insoluble forms. to avoid inclusion body formation, expression was optimized at 25 degrees c. furthermore, a 50% increase in solubilization of the target protein was obtained by adding 0.1 mm mg2+ to the medium. the purified recombinant gloshedobin gave a 44 ... | 2003 | 12882153 |
| expression of gloshedobin, a thrombin-like enzyme from the venom of gloydius shedaoensis, in escherichia coli. | gloshedobin, a thrombin-like enzyme from the venom of gloydius shedaoensis, was expressed in escherichia coli using expression vector pet-32a(+). the gene was expressed under t7 promotor with a fusion partner of thx.tag and a 6xhis.tag at its 5' terminal. after induction by iptg for 6 h, the recombinant enzyme was expressed in the cytoplasm. expression at 25 degrees c gave twice the amount of recombinant gloshedobin in cytoplasm than at 37 degrees c. | 2003 | 12882282 |
| purification, gene cloning and expression of an acidic phospholipase a2 from agkistrodon shedaoensis zhao. | a protein with the activity of phospholipase a(2) named asapla(2) was purified to homogeneity from the venom of agkistrodon shedaoensis zhao through deae-sepharose cl-6b anion exchange column, source s, and mono q fplc. its molecular weight was estimated to be 19 kd by sds-page, and its pi was about 3.5 by ief analysis. it inhibited the platelet aggregation that was induced by 1 micromol/ l adp, and the ic(50) was determined to be 6 micromol/l. degenerating primer was designed and synthesized ac ... | 2004 | 14732871 |
| chaperone-assisted column refolding of gloshedobin with the use of refolding cocktail. | gloshedobin, a recently isolated thrombin-like enzyme from the snake venom of gloydius shedaoensis, is expressed mainly in the form of inclusion bodies (ibs) in escherichia coli due to its cysteine-rich nature. following extraction and solubilization of the ibs, one-step immobilized metal affinity chromatography purification produces highly purified (>99%) denatured-solubilized gloshedobin ready to enter the subsequent refolding process. however, the traditional dilution or column refolding stra ... | 2008 | 19004452 |
| expression, purification and characterization of gloydius shedaoensis venom gloshedobin as hsp70 fusion protein in pichia pastoris. | gloshedobin, a thrombin-like enzyme from the venom of gloydius shedaoensis was expressed as hsp70 fusion protein from the construct ppic9k/hsp70-tle in the yeast pichia pastoris. by fusing gloshedobin to the c-terminus of hsp70, an expression level of 44.5mg hsp70-gloshedobin per liter of culture was achieved by methanol induction. the fusion protein secreted in the culture medium was conveniently purified by two chromatographic steps: q-sepharose ff and superdex 200. the purified enzyme had an ... | 2009 | 19286459 |
| soluble expression, purification, and characterization of gloydius shedaoensis venom gloshedobin in escherichia coli by using fusion partners. | gloshedobin, a thrombin-like enzyme from the venom of gloydius shedaoensis, is usually produced as inclusion bodies in escherichia coli cell. in this work, gloshedobin was separately fused with three fusion partners nusa, gst, and trxa at its n terminus and then was expressed as fusion proteins in e. coli. the results showed that the nusa was the most efficient fusion partner to improve the solubility of recombinant gloshedobin. the purified nusa-fused gloshedobin with an overall yield of 64.6% ... | 2010 | 19639313 |
| molecular mechanism analysis of gloydius shedaoensis venom gloshedobin interaction with inhibitors by homology modeling. | snake venom thrombin-like enzymes (svtles) are widely applied in the treatment of thrombotic diseases, however, the molecular mechanism of its inhibition by synthetic and natural proteinaceous inhibitors is not yet understood. here we investigated effects of protease inhibitors including phenylmethylsulfonil fluoride (pmsf), benzamidine (bmd) and its derivates on the activity of recombinant gloshedobin, a svtle from the snake gloydius shedaoensis. the molecular inhibition mechanism was postulate ... | 2010 | 20969888 |
| sequence and bioinformatic characterization of expressed sequence tags originated from gloydius shedaoensis shedaoensis venom gland. | first, a cdna library from gloydius shedaoensis shedaoensis venom gland (gssg) was constructed using smart technique. the total rna in a pair of gssgs was separated and the mrna was further isolated from it. the first-strand cdna was synthesized through powerscript(™) reverse transcriptase by a cds iii/3' pcr primer. a smart iv(™) -oligonucleotide was used as the template for the synthesis of double-strand cdna and amplified by long-distance polymerase chain reaction (ld-pcr). ld-pcr product was ... | 2013 | 23408674 |
| [phylogenetic relationships among viperidae, crotalinae based on mitochondrial 12s rrna sequence variations]. | this paper analyzed the phylogenetic relationships and classification of pit vipers (viperidae, crotalinae) based on mitochondrial 12s rrna gene sequence variations. we have sequenced mtdna 12s rrna gene about 370 bp fragment from gloydius saxatilis emelianov, gloydius shedaoensis zhao, gloydius ussurriensis (emelianov), trimeresurus stejnegeri schmidt and deinagkistrodon acutus, gloydius brevicaudus (stejneger) from two different localities, respectively. combined with the sequence of dinodon s ... | 2000 | 11147345 |
| the complete mitochondrial genome sequence of gloydius shedaoensis (squamata: viperidae). | gloydius shedaoensis is an insular and vulnerable pitviper that is endemic to snake island, northeastern china. in this study, we successfully sequenced mitochondrial genomes of two individuals of g. shedaoensis. the complete mitochondrial genomes of g. shedaoensis are circular molecular with 17 222 and 17 221 bp in length respectively, which both contain 2 ribosomal rna (rrna) genes, 13 protein-coding genes, 22 transfer rna (trna) genes, an origin of light-strand replication (ol) and two non-co ... | 2016 | 26644142 |
| "anatomical" view of the protein composition and protein characteristics for gloydius shedaoensis snake venom via proteomics approach. | the bioactive protein components from snake venom complexes have been utilized for studies of enzymology, structural biology, and pharmacology. the gloydius shedaoensis snake (gss) is the only snake species found exclusively at the chinese shedao (snake) island in dalian. to investigate the protein components of chinese gss venom (gssv), we initialized a proteomic assay for gssv by the combination of sodium dodecyl sulfate-polyacrylamide gel electrophoresis with high-performance liquid chromatog ... | 2011 | 21235002 |
| genetic adaptations of an island pit-viper to a unique sedentary life with extreme seasonal food availability. | the shedao pit-viper (gloydius shedaoensis) exhibits an extreme sedentary lifestyle. the island species exclusively feeds on migratory birds during migratory seasons and experiences prolonged hibernation and aestivation period each year (up to eight months). the sedentary strategy reduces energy expenditure, but may trigger a series of adverse effects and the snakes have likely evolved genetic modifications to alleviate these effects. to investigate the genetic adaptations, we sequenced and comp ... | 2020 | 32184370 |