Trypanosoma evansi: Cloning and Expression in Spodoptera fugiperda Insect Cells of the Diagnostic Antigen RoTat1.2

2001 ◽  
Vol 99 (4) ◽  
pp. 181-189 ◽  
Author(s):  
Toyohiko Urakawa ◽  
Didier Verloo ◽  
Luc Moens ◽  
Philippe Büscher ◽  
Phelix A.O. Majiwa
Keyword(s):  
Insect Cells ◽  
Trypanosoma Evansi ◽  
Cloning And Expression ◽  
Diagnostic Antigen ◽  
Spodoptera Fugiperda

Cloning and expression of HIV-1 p24 gene in insect cells by using BAC-TO-BAC system

1998 ◽  
Vol 3 (1) ◽  
pp. 113-118
Author(s):  
Mallam Nock Joshua ◽  
Qi Yipeng ◽  
Huang Yongxiu ◽  
Liu Ziye
Keyword(s):  
Insect Cells ◽  
Cloning And Expression ◽  
Hiv 1

scholarly journals Cytotoxic Cyplasin of the Sea Hare, Aplysia punctata, cDNA Cloning, and Expression of Bioactive Recombinants in Insect Cells

Neoplasia ◽  
2002 ◽  
Vol 4 (1) ◽  
pp. 49-59 ◽  
Author(s):  
Christian Petzelt ◽  
Gaby Joswig ◽  
Hermann Stammer ◽  
Dieter Werner
Keyword(s):  
Cdna Cloning ◽  
Insect Cells ◽  
Cloning And Expression ◽  
Sea Hare

Molecular cloning and expression in insect cells of honeybee venom allergen acid phosphatase (Api m 3)

2006 ◽  
Vol 117 (4) ◽  
pp. 848-854 ◽  
Author(s):  
T GRUNWALD ◽  
B BOCKISCH ◽  
E SPILLNER ◽  
J RING ◽  
R BREDEHORST ◽  
...  
Keyword(s):  
Acid Phosphatase ◽  
Molecular Cloning ◽  
Insect Cells ◽  
Cloning And Expression ◽  
Honeybee Venom ◽  
Venom Allergen

Tandem repeat protein as potential diagnostic antigen for Trypanosoma evansi infection

2011 ◽  
Vol 110 (2) ◽  
pp. 733-739 ◽  
Author(s):  
Nguyen Thu Thuy ◽  
Yasuyuki Goto ◽  
Zhao-Rong Lun ◽  
Shin-Ichiro Kawazu ◽  
Noboru Inoue
Keyword(s):  
Tandem Repeat ◽  
Trypanosoma Evansi ◽  
Repeat Protein ◽  
Diagnostic Antigen

Cloning and Expression in Insect Cells of two Pancreatic Lipases and a Procolipase from Myocastor coypus

1995 ◽  
Vol 227 (1-2) ◽  
pp. 186-193 ◽  
Author(s):  
Kenneth Thirstrup ◽  
Frederic Carriere ◽  
Siv A. Hjorth ◽  
Poul B. Rasmussen ◽  
Per F. Nielsen ◽  
...  
Keyword(s):  
Insect Cells ◽  
Cloning And Expression ◽  
Myocastor Coypus

scholarly journals Cloning and Expression of Mitochondrial Heat Shock Protein 70 of Trypanosoma congolense and Potential Use as a Diagnostic Antigen

2003 ◽  
Vol 10 (5) ◽  
pp. 926-933 ◽  
Author(s):  
Hiroshi Bannai ◽  
Tatsuya Sakurai ◽  
Noboru Inoue ◽  
Chihiro Sugimoto ◽  
Ikuo Igarashi
Keyword(s):  
Amino Acids ◽  
Heat Shock ◽  
Heat Shock Protein ◽  
Cdna Clone ◽  
Heat Shock Protein 70 ◽  
Trypanosoma Congolense ◽  
Cloning And Expression ◽  
Antibody Detection ◽  
Diagnostic Antigen ◽  
Mitochondrial Heat Shock Protein

ABSTRACT The ability to use mitochondrial heat shock protein 70 (MTP) of Trypanosoma congolense as a diagnostic antigen was examined. One cDNA clone was obtained by immunoscreening of a T. congolense procyclic form (PCF) cDNA library with monoclonal antibody (MAb) 10F9. The cDNA clone contained an open reading frame of 1,977 bp encoding a polypeptide consisting of 659 amino acids. Southern blotting analysis indicated that there were at least three copies of the MTP gene in the haploid genome. Interference of the MTP RNA resulted in complete inhibition, which indicated that MTP is essential at the PCF stage. Northern and Western blotting analyses revealed that MTP is expressed both in the bloodstream form (BSF) and in PCF. The B-cell epitope recognized by MAb 10F9 was located within 206 amino acids from the C terminus. Depending on the conditions of protein extraction, MTP was cleaved into smaller polypeptides by endogenous proteases. However, the C-terminal epitope of MTP was preserved with a high degree of antigenicity, even after cleavage. Antibody detection by enzyme-linked immunosorbent assay with the truncated recombinant MTP revealed that anti-MTP antibodies exist in experimentally infected mouse sera. Thus, MTP may be useful as an antigen for the serodiagnosis of primary T. congolense infection.

scholarly journals ERp60 does not substitute for protein disulphide isomerase as the β-subunit of prolyl 4-hydroxylase

1996 ◽  
Vol 316 (2) ◽  
pp. 599-605 ◽  
Author(s):  
Peppi KOIVUNEN ◽  
Tarja HELAAKOSKI ◽  
Pia ANNUNEN ◽  
Johanna VEIJOLA ◽  
Seija RÄISÄNEN ◽  
...  
Keyword(s):  
Amino Acid Sequence Identity ◽  
Insect Cells ◽  
Terminal Region ◽  
Cloning And Expression ◽  
Β Subunit ◽  
Hydroxylase Activity ◽  
Α Subunit ◽  
Protein Disulphide Isomerase ◽  
Tetramer Formation ◽  
Almost All

Prolyl 4-hydroxylase (EC 1.14.11.2) catalyses the formation of 4-hydroxyproline in collagens. The vertebrate enzymes are α2β2 tetramers while the Caenorhabditis elegans enzyme is an αβ dimer. The β-subunit is identical to protein disulphide isomerase (PDI), a multifunctional endoplasmic reticulum luminal polypeptide. ERp60 is a PDI isoform that was initially misidentified as a phosphatidylinositol-specific phospholipase C. We report here on the cloning and expression of the human and Drosophila ERp60 polypeptides. The overall amino acid sequence identity and similarity between the processed human ERp60 and PDI polypeptides are 29% and 56% respectively, and those between the Drosophila ERp60 and human PDI polypeptides 29% and 55%. The two ERp60 polypeptides were found to be similar to human PDI within almost all their domains, the only exception being the extreme C-terminal region. Nevertheless, when the human or Drosophila ERp60 was expressed in insect cells together with an α-subunit of human prolyl 4-hydroxylase, no tetramer was formed and no prolyl 4-hydroxylase activity was generated in the cells. Additional experiments with hybrid polypeptides in which the C-terminal regions had been exchanged between the human ERp60 and PDI polypeptides demonstrated that the differences in the C-terminal region are not the only reason for the lack of prolyl 4-hydroxylase tetramer formation by ERp60.

cDNA cloning and expression of a gamma-aminobutyric acidA receptor epsilon-subunit in rat brain

2000 ◽  
Vol 12 (12) ◽  
pp. 4318-4330 ◽  
Author(s):  
Nathalie Moragues ◽  
Philippe Ciofi ◽  
Pierrette Lafon ◽  
Marie-Francoise Odessa ◽  
Gerard Tramu ◽  
...  
Keyword(s):  
Rat Brain ◽  
Cdna Cloning ◽  
Cloning And Expression ◽  
Epsilon Subunit

Cloning and Expression of Recombinant Human Insulin Gene in Pichia pastoris

2019 ◽  
Vol 10 (01) ◽  
Author(s):  
Rafid A. Abdulkareem
Keyword(s):  
Pichia Pastoris ◽  
Human Insulin ◽  
Expression Vector ◽  
Expression System ◽  
Insulin Gene ◽  
Cloning And Expression ◽  
Pcr Analysis ◽  
Major Band ◽  
Human Insulin Gene ◽  
Pichia Pastoris Expression System

The main goal of the current study was cloning and expression of the human insulin gene in Pichia pastoris expression system, using genetic engineering techniques and its treatment application. Total RNA was purified from fresh normal human pancreatic tissue. RNA of good quality was chosen to obtain a first single strand cDNA. Human preproinsulin gene was amplified from cDNA strand, by using two sets of specific primers contain EcoR1 and Notl restriction sites. The amplified preproinsulin gene fragment was double digested with EcoRI and Not 1 restriction enzymes, then inserted into pPIC9K expression vector. The new pPIC9K-hpi constructive expression vector was transformed by the heat-shock method into the E.coli DH5α competent cells. pPic9k –hpi, which was propagated in the positive transformant E. coli cells, was isolated from cells and then linearised by restriction enzyme SalI, then transformed into Pichia pastoris GS115 using electroporation method. Genomic DNA of His+ transformants cell was extracted and used as a template for PCR analysis. The results showed, that the pPic9k – hpi was successfully integrated into the P. pastoris genome, for selected His+ transformants clones on the anticipated band at 330 bp, which is corresponded to the theoretical molecular size of the human insulin gene. To follow the insulin expression in transformans, Tricine–SDS gel electrophoresis and Western blot analysis were conducted. The results showed a successful expression of recombinant protein was detected by the presence of a single major band with about (5.8 KDa) on the gel. These bands correspond well with the size of human insulin with the theoretical molecular weight (5.8 KDa).

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