Abstract:
AIM:To study the influence of redox environment of Escherichia coli (E. coli) cytoplasm on disulfide bond formation of recombinant proteins. METHODS:Bovine fibroblast growth factor (BbFGF) was selected as a model of simple proteins with a single disulfide bond and free cysteines. Anti-HBsAg single-chain Fv (HBscFv), an artificial multidomain protein, was selected as the model molecule of complex protein with 2 disulfide bonds. A BbFGF-producing plasmid, pJN-BbFGF, and a HBscFv producing-plasmid, pQE-HBscFv, were constructed and transformed into E. coli strains BL21(DE3) and M15(pREP4) respectively. At the same time, both plasmids were transformed into a reductase-deficient host strain, E. coli Origami(DE3). The 4 recombinant E. coli strains were cultured and the target proteins were purified. Solubility and bioactivity of recombinant BbFGF and HBscFv produced in different host strains were analyzed and compared respectively. RESULTS:All recombinant E. coli strains could efficiently produce target proteins. The level of BbFGF in BL21(DE3) was 15-23% of the total protein, and was 5-10% in Origami (DE3). In addition, 65% of the BbFGF produced in BL21(DE3) formed into inclusion body in the cytoplasm, and all the target proteins became soluble in Origami(DE3). The bioactivity of BbFGF purified from Origami(DE3) was higher than its counterpart from BL21(DE3). The ED(50) of BbFGF from Origami(DE3) and BL21(DE3) was 1.6 microg/L and 2.2 microg/L, respectively. Both HBscFv formed into inclusion body in the cytoplasm of M15(pQE-HBscFv) or Origami(pQE-HBscFv). But the supernatant of Origami(pQE-HBscFv) lysate displayed weak bioactivity and its counterpart from M15(pQE-HBscFv) did not display any bioactivity. The soluble HBscFv in Origami(pQE-HBscFv) was purified to be 1-2 mg/L and its affinity constant was determined to be 2.62 x 10(7) mol/L. The yield of native HBscFv refolded from inclusion body in M15(pQE-HBscFv) was 30-35 mg/L and the affinity constant was 1.98 x 10(7) mol/L. There was no significant difference between the bioactivity of HBscFvs refolded from the inclusion bodies produced in different host strains. CONCLUSION:Modification of the redox environment of E. coli cytoplasm can significantly improve the folding of recombinant disulfide-bonded proteins produced in it.
journal_name
World J Gastroenteroljournal_title
World journal of gastroenterologyauthors
Xiong S,Wang YF,Ren XR,Li B,Zhang MY,Luo Y,Zhang L,Xie QL,Su KYdoi
10.3748/wjg.v11.i7.1077keywords:
subject
Has Abstractpub_date
2005-02-21 00:00:00pages
1077-82issue
7eissn
1007-9327issn
2219-2840journal_volume
11pub_type
杂志文章abstract::The advances of laparoscopic surgery since the early 1990s have caused one of the largest technical revolutions in medicine since the detection of antibiotics (1922, Flemming), the discovery of DNA structure (1953, Watson and Crick), and solid organ transplantation (1954, Murray). Perseverance through a rocky start an...
journal_title:World journal of gastroenterology
pub_type: 杂志文章,评审
doi:10.3748/wjg.v20.i41.15119
更新日期:2014-11-07 00:00:00
abstract:AIM:To measure the normal space between the posterior wall of the vagina and the anterior wall of the respectively rectum using computed tomography (CT) and reveal its were relationship to rectocele. METHODS:A total of twenty female volunteers without rectocele were examined by CT scan. We performed a middle level con...
journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.15.3051
更新日期:2009-06-28 00:00:00
abstract:AIM:Both Hepatitis B virus (HBV) and Hepatitis C virus (HCV) are major causative agents of transfusion-associated and community-acquired hepatitis worldwide. Development of a HCV vaccine as well as more effective HBV vaccines is an urgent task. DNA immunization provides a promising approach to elicit protective humoral...
journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v8.i3.505
更新日期:2002-06-01 00:00:00
abstract:AIM:To validate the Rockall scoring system for predicting outcomes of rebleeding, and the need for a surgical procedure and death. METHODS:We used data extracted from the Registry of Upper Gastrointestinal Bleeding and Endoscopy including information of 1869 patients with non-variceal upper gastrointestinal bleeding t...
journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v12.i48.7779
更新日期:2006-12-28 00:00:00
abstract::Alcohol-induced fatty liver (steatosis) was believed to result from excessive generation of reducing equivalents from ethanol metabolism, thereby enhancing fat accumulation. Recent findings have revealed a more complex picture in which ethanol oxidation is still required, but specific transcription as well as humoral ...
journal_title:World journal of gastroenterology
pub_type: 杂志文章,评审
doi:10.3748/wjg.v13.i37.4974
更新日期:2007-10-07 00:00:00
abstract::Hepatocellular carcinoma (HCC) is the most common primary liver cancer with a dismal prognosis, especially when diagnosed at advanced stages. Annexin A2 (ANXA2), is found to promote cancer progression and therapeutic resistance. However, the underlining mechanisms of ANXA2 in immune escape of HCC remain poorly underst...
journal_title:World journal of gastroenterology
pub_type: 杂志文章,评审
doi:10.3748/wjg.v26.i18.2126
更新日期:2020-05-14 00:00:00
abstract:AIM:To determine the expression levels of gastrointestinal nesfatin-1 in ventromedial hypothalamic nucleus (VMH)-lesioned (obese) and ventrolateral hypothalamic nucleus (VLH)-lesioned (lean) rats that exhibit an imbalance in their energy metabolism and gastric mobility. METHODS:Male Wistar rats were randomly divided i...
journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v20.i22.6897
更新日期:2014-06-14 00:00:00
abstract:AIM:Because of a major resistance to chemotherapy, prognosis of hepatocellular carcinoma (HCC) is still poor. New treatments are required and gene therapy may be an option. Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) induces apoptosis in multiple malignant tumors, and using adenoviral vectors has sh...
journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v11.i17.2552
更新日期:2005-05-07 00:00:00
abstract:AIM:To investigate the prognostic significance of PECAM-1, ICAM-3 and HLA-DR antigens in patients with primary non-Hodgkin's gastric lymphoma. METHODS:We immunohistochemically studied PECAM-1, ICAM-3 and HLA-DR antigen expression in 36 B-cell MALT-type primary gastric lymphoma patients. Ten non-malignant and ten healt...
journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v12.i12.1924
更新日期:2006-03-28 00:00:00
abstract:AIM:To determine whether intra-arterial infusion of triolein emulsion has biochemical and histopathologic effect on rabbit liver. METHODS:An emulsion of 0.2 mL triolein in 20 mL of saline was infused into either the hepatic arteries of nine rabbits (group 1) or the superior mesenteric arteries of 12 rabbits (group 2)....
journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v20.i39.14442
更新日期:2014-10-21 00:00:00
abstract:AIM:To report the clinical outcomes and ergonomics analysis of three laparoscopic approaches in the management of Hirschsprung's disease (HD). METHODS:There were 90 pediatric patients (63 boys, 27 girls; mean age: 3.6 ± 2.7 mo; range: 1.0-90.2 mo) who underwent laparoscopic endorectal pull-through Soave procedures for...
journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v21.i29.8903
更新日期:2015-08-07 00:00:00
abstract:AIM:To characterize the protein files in blood from same patients with esophageal squamous cell carcinoma (ESCC) before and after operation at the high-incidence area for ESCC in Henan Province, China. METHODS:Two-dimensional electrophoresis, silver staining and ImageMaster 2-DE analysis software were applied to the d...
journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v10.i22.3365
更新日期:2004-11-15 00:00:00
abstract:AIM:To investigate the changes that occur in E-cadherin expression during the process of metastasis in colorectal cancer. METHODS:E-cadherin expression was detected by immunohistochemistry and two indices of expression were calculated which reflected the level of expression and the locations (membrane and cytoplasm). ...
journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v12.i27.4304
更新日期:2006-07-21 00:00:00
abstract::Colonic motility subserves large bowel functions, including absorption, storage, propulsion and defaecation. Colonic motor dysfunction remains the leading hypothesis to explain symptom generation in chronic constipation, a heterogeneous condition which is extremely prevalent in the general population, and has huge soc...
journal_title:World journal of gastroenterology
pub_type: 社论,评审
doi:10.3748/wjg.v16.i41.5162
更新日期:2010-11-07 00:00:00
abstract::Somatostatinomas are extremely rare neuroendocrine tumors of the gastrointestinal tract, first described in the pancreas in 1977 and in the duodenum in 1979. They may be functional and cause somatostatinoma or inhibitory syndrome, but more frequently are non-functioning pancreatic endocrine tumors that produce somatos...
journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.15.5859
更新日期:2009-12-14 00:00:00
abstract:AIM:To determine the clinical effectiveness of intense psychological support to physical activity (PA) in nonalcoholic fatty liver disease (NAFLD), compared with cognitive-behavioral treatment (CBT). METHODS:Twenty-two NAFLD cases received support to exercise, tailored to their motivational needs (PA group). The effec...
journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v20.i29.10128
更新日期:2014-08-07 00:00:00
abstract:AIM:To clarify the effect of a high des-gamma-carboxy prothrombin (DCP) level on the invasiveness and prognosis of small hepatocellular carcinoma. METHODS:Among 142 consecutive patients with known DCP levels, who underwent hepatectomy because of hepatocellular carcinoma, 85 patients met the criteria for small hepatoce...
journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.14.1370
更新日期:2008-03-07 00:00:00
abstract:AIM:To investigate the diagnosis of nonalcoholic steatohepatitis (NASH) using contrast ultrasonography in the NASH rat model. METHODS:The liver in methionine choline-deficient diet (MCDD) rats, a NASH model constructed by feeding an MCDD, was examined by contrast ultrasonography at weeks 2, 4, 8, 12 and 16, with late ...
journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v17.i37.4191
更新日期:2011-10-07 00:00:00
abstract:AIM:Hepatocellular carcinoma (HCC) is a common disease in Taiwan. The prevalence of viral hepatitis infection and the subsequent development of HCC are well known to be higher in patients with end-stage renal disease (ESRD) requiring hemodialysis (HD) or peritoneal dialysis (PD) than among the general population. Howev...
journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v11.i14.20
更新日期:2005-04-14 00:00:00
abstract::It is one of the most important problems for general surgeons to decide which operation should be undertaken on patients with intra-abdominal infection, especially those with concomitant abdominal hypertension. Recently, closure techniques using prosthetic meshes in order to retain abdominal tension and to control sep...
journal_title:World journal of gastroenterology
pub_type: 信件
doi:10.3748/wjg.14.815
更新日期:2008-02-07 00:00:00
abstract:AIM:To evaluate the clinical outcomes of patients undergoing hepatectomy with hemihepatic vascular occlusion (HHO) compared with total hepatic inflow occlusion (THO). METHODS:Randomized controlled trials (RCTs) comparing hemihepatic vascular occlusion and total hepatic inflow occlusion were included by a systematic li...
journal_title:World journal of gastroenterology
pub_type: 杂志文章,meta分析,评审
doi:10.3748/wjg.v17.i26.3158
更新日期:2011-07-14 00:00:00
abstract:AIM:To investigate the effects of gastric pacing on gastric emptying and plasma motilin level in a canine model of gastric motility disorders and the correlation between gastric emptying and plasma motilin level. METHODS:Ten healthy Mongrel dogs were divided into: experimental group of six dogs and control group of fo...
journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v10.i3.419
更新日期:2004-02-01 00:00:00
abstract:AIM:To assess the diagnostic accuracy of endoscopic ultrasound (EUS), fluid tumor markers and cytology in distinguishing benign from (pre)malignant pancreatic cystic lesions. METHODS:46 consecutive patients, referred to a gastroenterologist and surgeon for a symptomatic or incidental pancreatic cyst, were reviewed. EU...
journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v13.i29.3962
更新日期:2007-08-07 00:00:00
abstract:AIM:To prospectively explore the role of carcinoembryonic antigen (CEA) in gallbladder bile in patients with colorectal carcinoma and the morphological and clinical features of neoplasia and the occurrence of hepatic metastases. METHODS:CEA levels in the gallbladder and peripheral blood were studied in 44 patients wit...
journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v9.i7.1589
更新日期:2003-07-01 00:00:00
abstract::Folate deficiency and its association with cancer have been studied in the literature, but its clinical impact is still unknown. Folate deficiency and its result on gastric cancer is a mysterious part of oncology, with ongoing studies hopefully clarifying its impact on gastric cancer management. Lee et al studied fola...
journal_title:World journal of gastroenterology
pub_type: 评论,信件
doi:10.3748/wjg.v21.i13.4101
更新日期:2015-04-07 00:00:00
abstract::Sorafenib has been considered the standard of care for patients with advanced unresectable hepatocellular carcinoma (HCC) since 2007 and numerous studies have investigated the role of markers involved in the angiogenesis process at both the expression and genetic level and clinical aspect. What results have ten years ...
journal_title:World journal of gastroenterology
pub_type: 杂志文章,评审
doi:10.3748/wjg.v24.i36.4152
更新日期:2018-09-28 00:00:00
abstract:AIM:To investigate the gastrin secreting cells (G cells) and the somatostatin secreting cells (D cells) of antral mucosa in rats at the ultrastructural level. METHODS:Revised immunoelectron microscopic technique was used to detect the G cells and D cells in rat antral mucosa through gastrin and somatostatin antibodies...
journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v9.i12.2768
更新日期:2003-12-01 00:00:00
abstract::Liver cancer is one of the world's most common cancers and the second leading cause of cancer deaths. Hepatocellular carcinoma (HCC), a primary hepatic cancer, accounts for 90%-95% of liver cancer cases. The pathogenesis of HCC consists of a stepwise process of liver damage that extends over decades, due to hepatitis,...
journal_title:World journal of gastroenterology
pub_type: 杂志文章,评审
doi:10.3748/wjg.v21.i42.12042
更新日期:2015-11-14 00:00:00
abstract:AIM:To describe the significant over-expression of fibroblast growth factor receptor 3 (FGFR3), which is a signal transduction and cell proliferation related gene in hepatocellular carcinoma (HCC). METHODS:Following DNA microarray, Northern blot and quantitative real-time PCR were employed to confirm FGFR3 expression ...
journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v11.i34.5266
更新日期:2005-09-14 00:00:00
abstract:AIM:To investigate whether single endoscopist-performed percutaneous endoscopic gastrostomy (PEG) is safe and to compare the complications of PEG with those reported in the literature. METHODS:Patients who underwent PEG placement between June 2001 and August 2011 at the Baskent University Alanya Teaching and Research ...
journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v19.i26.4172
更新日期:2013-07-14 00:00:00