Contact Transfer Method: DNA Transfer from Polyacrylamide Gel

1995 ◽  
Vol 224 (2) ◽  
pp. 611-612 ◽  
Author(s):  
M. Takeuchi ◽  
H. Fujisawa
Keyword(s):  
Polyacrylamide Gel ◽  
Dna Transfer ◽  
Transfer Method

Efficient and simple production of transgenic mice and rabbits using the new DMSO-sperm mediated exogenous DNA transfer method

2006 ◽  
Vol 73 (5) ◽  
pp. 589-594 ◽  
Author(s):  
Wei Shen ◽  
Lan Li ◽  
Qingjie Pan ◽  
Lingjiang Min ◽  
Huansheng Dong ◽  
...  
Keyword(s):  
Transgenic Mice ◽  
Dna Transfer ◽  
Exogenous Dna ◽  
Transfer Method ◽  
Simple Production

scholarly journals Highly Efficient CRISPR/Cas9 System in Plasmodium Falciparum Using Cas9-expressing Parasites and a Linear Donor Template

2021 ◽  
Author(s):  
Tsubasa Nishi ◽  
Naoaki Shinzawa ◽  
Masao Yuda ◽  
Shiroh Iwanaga
Keyword(s):  
Plasmodium Falciparum ◽  
High Efficiency ◽  
Standard Technique ◽  
Dna Transfer ◽  
Drug Selection ◽  
Technical Problems ◽  
Highly Efficient ◽  
Transgenic Parasites ◽  
Transfer Method ◽  
Donor Template

Abstract The current CRISPR/Cas9 system for Plasmodium falciparum suffers from technical problems caused by plasmid constructs, such as delays in establishing transgenic parasites during drug selection and unexpected integration of circular donor DNA by single-crossover recombination. Although these problems can be solved by using linear donor templates, such an approach requires highly efficient introduction of DNA and rapid completion of recombination because linear DNA is easily lost from the parasites during multiplication. Here, we overcame these problems by developing a highly efficient DNA transfer method and Cas9-expressing parasites. Using our new CRISPR/Cas9 system, transgenic parasites were established in two weeks without any unexpected recombination or off-target mutations. Furthermore, with our system, two genes on different chromosomes were successfully modified in one transfection. Because of its high efficiency and robustness, our new CRISPR/Cas9 system will become a standard technique for genetic engineering of P. falciparum and dramatically advance studies of this parasite.

scholarly journals Production of Transgenic Goats by Sperm-mediated Exogenous DNA Transfer Method

2009 ◽  
Vol 23 (1) ◽  
pp. 33-40 ◽  
Author(s):  
Yongju Zhao ◽  
Hong Wei ◽  
Yong Wang ◽  
Lingbin Wang ◽  
Mingju Yu ◽  
...  
Keyword(s):  
Dna Transfer ◽  
Exogenous Dna ◽  
Transgenic Goats ◽  
Transfer Method

Ultrastructural localization of specific nucleoprotein fractions

1978 ◽  
Vol 36 (2) ◽  
pp. 204-205
Author(s):  
G. L. Brown
Keyword(s):  
Gel Electrophoresis ◽  
Mouse Liver ◽  
Polyacrylamide Gel Electrophoresis ◽  
Ultrastructural Localization ◽  
Polyacrylamide Gel ◽  
Acid Extraction ◽  
Acid Solutions ◽  
Low Salt ◽  
Liver Nuclei ◽  
Phosphate Groups

Bismuth (Bi) stains nucleoproteins (NPs) by interacting with available amino and primary phosphate groups. These two staining mechanisms are distinguishable by glutaraldehyde crosslinking (Fig. 1,2).Isolated mouse liver nuclei, extracted with salt and acid solutions, fixed in either formaldehyde (form.) or gl utaraldehyde (glut.) and stained with Bi, were viewed to determine the effect of the extractions on Bi stainina. Solubilized NPs were analyzed by SDS-polyacrylamide gel electrophoresis.Extraction with 0.14 M salt does not change the Bi staining characteristics (Fig. 3). 0.34 M salt reduces nucleolar (Nu) staining but has no effect on interchromatinic (IC) staining (Fig. 4). Proteins responsible for Nu and glut.- insensitive IC staining are removed when nuclei are extracted with 0.6 M salt (Fig. 5, 6). Low salt and acid extraction prevents Bi-Nu staining but has no effect on IC staining (Fig. 7). When nuclei are extracted with 0.6 M salt followed by low salt and acid, all Bi-staining components are removed (Fig. 8).

The Effect of Antithrombin III on the Activity of the Coagulation Factors VII, IX and X

1976 ◽  
Vol 35 (02) ◽  
pp. 295-304 ◽  
Author(s):  
B Østerud ◽  
M Miller-Andersson ◽  
U Abildgaard ◽  
H Prydz
Keyword(s):  
Antithrombin Iii ◽  
Factor Xa ◽  
Coagulation Factors ◽  
Disc Electrophoresis ◽  
Polyacrylamide Gel ◽  
Factor Vii ◽  
Native Form ◽  
Factor Ixa ◽  
Plasma Factor

SummaryAntithrombin III, purified to homogeneity according to Polyacrylamide gel disc electrophoresis and immunoelectrophoresis, inhibited the activity of purified factor IXa and Xa, whereas factor VII was not inhibited either in the active or in the native form.Antithrombin III is the single most important inhibitor of factor Xa in plasma. Factor Xa does not, however, reduce the activity of antithrombin III against thrombin.

Fibrinogen Montreal

1973 ◽  
Vol 29 (03) ◽  
pp. 536-546 ◽  
Author(s):  
M Lacombe ◽  
J Soria ◽  
C Soria ◽  
G d’Angelo ◽  
R Lavallee ◽  
...  
Keyword(s):  
Optical Density ◽  
Polyacrylamide Gel ◽  
Clotting Time ◽  
Functional Defect ◽  
A Chain ◽  
Fibrin Monomers

SummaryA new case of congenital dysfibrinogenemia characterized by a prolonged thrombin clotting time and a low optical density of the polymerization curve has been discovered in Montreal. The functional defect is due to an abnormal aggregation of fibrin monomers.The characteristics of this abnormal fibrinogen are serum gélification (Paracoagulation) at 37°, 22° and 4° C, a normal immuno-electrophoretic and electrofocusing pattern, a slight increase in the mobility in the α (A) chain by electrophoresis of the dissociated chains in polyacrylamide gel. However, no abnormality was found in the α (A) chain of the disulphide knot.

Platelet Microtubule Subunit Proteins

1979 ◽  
Vol 42 (05) ◽  
pp. 1630-1633 ◽  
Author(s):  
A G Castle ◽  
N Crawford
Keyword(s):  
Epithelial Cells ◽  
Gel Electrophoresis ◽  
Blood Platelets ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Lens Epithelial Cells ◽  
Molecular Weights ◽  
Kinetics Of ◽  
Microtubular Structures

SummaryBlood platelets contain microtubule proteins (tubulin and HMWs) which can be polymerised “in vitro” to form structures which resemble the microtubules seen in the intact platelet. Platelet tubulin is composed of two non-identical subunits a and p tubulin which have molecular weights around 55,000 but can be resolved in alkaline SDS-polyacrylamide gel electrophoresis. These subunits associate as dimers with sedimentation coefficients of about 5.7 S although it is not known whether the dimer protein is a homo- or hetero-dimer. The dimer tubulin binds the anti-mitotic drug colchicine and the kinetics of this binding are similar to those reported for neurotubulins. Platelet microtubules also contain two HMW proteins which appear to be essential and integral components of the fully assembled microtubule. These proteins have molecular weights greater than 200,000 daltons. Fluorescent labelled antibodies to platelet and brain tubulins stain long filamentous microtubular structures in bovine lens epithelial cells and this pattern of staining is prevented by exposing the cells to conditions known to cause depolymerisation of cell microtubules.

Sign in / Sign up

Export Citation Format

Share Document