scholarly journals Cloning and expression of the flavin reductase LuxG from Photobacterium leiognathi YL and its improvement for NADH detection

2020 ◽  
Vol 19 (2) ◽  
pp. 274-280 ◽  
Author(s):  
Guanhua Xuan ◽  
Qilin Xiao ◽  
Jingxue Wang ◽  
Hong Lin
Keyword(s):  
Cloning And Expression ◽  
Flavin Reductase ◽  
Photobacterium Leiognathi ◽  
Higher Sensitivity

LuxG from P. leiognathi YL was successfully purified and characterized. Based on this, a coupled pure enzyme bioluminescent system was established and used for NADH detection, which showed higher sensitivity than existing bioluminescent systems.

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

Species Specific Immunoassays to Measure Blood Platelet and Coagulation Activation in the Rat

1996 ◽  
Vol 76 (06) ◽  
pp. 1090-1095 ◽  
Author(s):  
C Ravanat ◽  
M Freund ◽  
S Schuhler ◽  
P Grunert ◽  
L Meyer ◽  
...  
Keyword(s):  
Antithrombin Iii ◽  
Plasma Concentrations ◽  
Simultaneous Detection ◽  
Blood Platelet ◽  
Coagulation Activation ◽  
Platelet Factor ◽  
Prethrombotic State ◽  
Low Levels ◽  
Species Specific ◽  
Higher Sensitivity

SummaryThe purpose of this study was to develop specific and sensitive immunoassays to detect early indices of hypercoagulability in the rat. Rat platelet factor 4 (rPF4) and rat fibrinopeptide A (rFPA) assays, tools for the detection of activation of platelets and coagulation respectively, were designed using antibodies raised against purified rPF4 and against synthetic rFPA. The relevance of these new assays and of the commercially available ELISA kit for thrombin-antithrombin III (TAT) complexes was demonstrated in a rat model of a prethrombotic state induced by intravenous infusion of varying doses of thromboplastin (90 to 2400 μl/kg/h). In this model, the immunoassays allowed simultaneous detection of low levels of rFPA and rPF4 which were correlated with fibrinogen and platelet consumption and TAT generation and further proved to be of higher sensitivity than the classical methods of platelet count or measurement of fibrinogen levels. Plasma concentrations of rFPA, rPF4 and TAT were dependent on infusion time and thromboplastin dose, while hirudin (1 mg/kg) prevented their appearance. Thus the new specific immunoassays for rPF4 and rFPA and the commercial human TAT assay represent useful tools for pathophysiological studies or the screening of antithrombotic drugs in rats.

Pituitary and ovarian response to transient doe-litter separation in nursing rabbits

Reproduction ◽  
2000 ◽  
pp. 361-366 ◽  
Author(s):  
E Ubilla ◽  
PG Rebollar ◽  
D Pazo ◽  
AI Esquifino ◽  
JM Alvarino
Keyword(s):  
Pituitary Gland ◽  
Litter Size ◽  
Artificial Insemination ◽  
Ovarian Response ◽  
Major Effect ◽  
Free Access ◽  
Plasma Prolactin ◽  
Follicular Waves ◽  
Higher Sensitivity

The effects of a transient doe-litter separation on plasma prolactin, FSH and oestradiol concentrations, as well as the effect on LH response to exogenous GnRH administered at the time of artificial insemination, were determined in nursing rabbits. The effects on fertility, and litter size after parturition, as well as litter survival after doe-litter separation, were also studied. Control does (n = 12) had free access to nursing, whereas biostimulated does (n = 12) were separated from their litters for 48 h before artificial insemination. Plasma prolactin concentrations were decreased 24 h after the doe-litter separation (P < 0.05). The response of prolactin to suckling reached 10 times the basal values measured on day 10 after parturition (P < 0.0001). Increased oestradiol concentrations were found during the 48 h after the doe-litter separation: at 0 h, before artificial insemination (P< 0. 0001), 1.0-2.0 h after artificial insemination (P < 0.001), at 2.5 h (P < 0.05), 3.0 h (P < 0.01), and at 3.5 h (P < 0.05) after artificial insemination. Exogenous GnRH administered at the time of artificial insemination caused a greater LH response in does previously separated from their litters during 48 h (P < 0.01). The transient doe-litter separation did not affect plasma FSH concentrations, fertility, litter size or litter survival. These results suggest that a transient separation of nursing does from their litters before artificial insemination results in a decrease in plasma prolactin concentrations that could promote growth of follicular waves, and high steroidogenesis activity, leading to increased oestradiol concentrations and inducing higher sensitivity of the pituitary gland to exogenous GnRH. These findings associated to the absence of suckling episodes would lead to higher LH response and, therefore, exert a major effect on fertility.

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).

APPLICATION OF SHIFTING ASSAY OF STANDARDIZED INDIRECT AGGLUTINATION (SSIA) FOR DETECTION OF ANTIGENS OF NEWCASTLE DISEASE AND INFECTIOUS BURSAL DISEASE VIRUSES IN CHICKEN FAECES

2013 ◽  
Vol 83 (5) ◽  
Author(s):  
Phạm Hồng Sơn ◽  
Phạm Hồng Kỳ ◽  
Nguyễn Thị Lan Hương ◽  
Phạm Thị Hồng Hà
Keyword(s):  
Newcastle Disease ◽  
Winter Season ◽  
Summer Season ◽  
Infectious Bursal Disease ◽  
Infection Rates ◽  
Faecal Samples ◽  
Infectious Bursal Disease Viruses ◽  
Bursal Disease ◽  
Long Time ◽  
Higher Sensitivity

. Using the method of shifting assay of standardized indirect agglutination (SSIA), the prevalence of Newcastle disease viruses (NDV) and infectious bursal disease viruses (IBDV) in chickens reared in several districts of Thua Thien Hue province in the Spring-Summer and Fall-Winter seasons was determined. In the Spring-Summer season of 2011, about 22.3% of the chickens were infected with NDV, in which A Luoi  accounted for the highest percentage of 25% of the infected chickens and Huong Thuy  the lowest  of 18.2%. Meanwhile, 36% of the same chickens were infected with IBDV, with the highest percentage (46.66%) also in A Luoi and the lowest (30.3%) also in Huong Thuy. The intensity of NDV infection in the Spring-Summer season in A Luoi and Phu Vang was highest (GMT = 1.45), and in Huong Thuy lowest (GMT = 1.31). In addition, in the Fall-Winter season, about 46% of the chickens were infected with NDV and 46.3% with IBDV in Huong Thuy and Phu Vang – two neighbouring districts of Hue City, in which NDV was detected in 54.4% of the chickens in Huong Thuy and 33.9% in Phu Vang. In contrast, IBDV was detected in 41.9% and 52.7% of the chickens respectively in the two districts. The infection was not inter-dependent. Methodically, although the differences in the infection rates were insignificant with the accuracy of 95%, faecal samples showed higher sensitivity in SSIA analyses for both cases of NDV and IBDV infection in comparision with mouth exudates. By SSIA method, results could be read clearly with unaided eyes for a long time after the performance, and it was also proven applicable for cases of haemagglutinating viruses if proper treatments for depletion of animal RBCs’ surface agglutinins could be applied.

Application of Scan Based FA and Advanced Passive Voltage Contrast Technique in Defect Isolation

2003 ◽  
Author(s):  
Michael B. Schmidt ◽  
Noor Jehan Saujauddin
Keyword(s):  
Failure Analysis ◽  
Large Scale ◽  
Fault Isolation ◽  
Electronic Information ◽  
Scan Testing ◽  
Challenging Environment ◽  
Contrast Technique ◽  
Defect Isolation ◽  
Voltage Contrast ◽  
Higher Sensitivity

Abstract Scan testing and passive voltage contrast (PVC) techniques have been widely used as failure analysis fault isolation tools. Scan diagnosis can narrow a failure to a given net and passive voltage contrast can give real-time, large-scale electronic information about a sample at various stages of deprocessing. In the highly competitive and challenging environment of today, failure analysis cycle time is very important. By combining scan FA with a much higher sensitivity passive voltage contrast technique, one can quickly find defects that have traditionally posed a great challenge.

INFLUENCE OF SODIUM THIOPENTAL ON CONSERVATION OF CONDITIONAL REFLEX OF PASSIVE AVOIDANCE IN RATS WITH DIFFERENT NERRVOUS SYSTEM EXCITABILITY

2018 ◽  
pp. 8-11
Author(s):  
A. V. Shvetsov ◽  
A. I. Vaido ◽  
N. A. Dyuzhikova ◽  
A. V. Belskaya ◽  
M. V. Mikhailova ◽  
...  
Keyword(s):  
Nervous System ◽  
Passive Avoidance ◽  
Avoidance Response ◽  
Conditioned Reflex ◽  
Lethal Dose ◽  
Conditional Reflex ◽  
Passive Avoidance Response ◽  
Sodium Thiopental ◽  
Higher Sensitivity

A study of the dynamics of preservation of the conditioned reflex of passive avoidance (passive avoidance response) in rats of two lines with different level of excitability of the nervous system was performed: with a high and low thresholds of excitability as normal and after exposure to sodium thiopental semi-lethal dose. It was shown that a long preservation of the memorable trace under standard conditions and a higher sensitivity to the sodium thiopental action manifest in rats with a high excitability threshold in comparison with low-excitable line of rats.

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