Efficient site directed in vitro mutagenesis using ampicillin selection potx
Efficient site directed in vitro mutagenesis using ampicillin selection potx
... Nucleic Acids Research, Vol. 18, No. 12 3439 Efficient site directed in vitro mutagenesis using ampicillin selection Martin K.Lewis* and David V.Thompson Promega Corporation, Madison, WI 5371 1, USA Received April 11, 1990; Accepted May 25, 1990 ABSTRACT A novel plasmid vector pSELECT-1 is described which can be used for highly efficient site- directed in vitro mut...
Ngày tải lên: 23/03/2014, 22:20
in vitro mutagenesis protocols, 2nd
... Combining Site- Specific Chemical Modification with Site- Directed Mutagenesis: Versatile Strategy to Move Beyond Structural Limitations of 20 Natural Amino Acids Side Chains in Protein Engineering Grace ... Deletions, and Insertions, Using QuikChange™ Site- Directed Mutagenesis Wenyan Wang and Bruce A. Malcolm 37 6 Efficient and Accurate Site- Directed Mutagenesis of L...
Ngày tải lên: 11/04/2014, 06:56
in vitro mutagenesis protocols
... Using the Unique Restriction Site Elimination (USE) Method Li Zhu 1. Introduction In vitro site- directed mutagenesis has been widely used in vector modi- fication, and in gene and protein ... restriction site located in an intergemc region as the selection site. If the selection restriction site must be located within a gene, avoid using a selection primer...
Ngày tải lên: 11/04/2014, 09:47
Tài liệu Báo cáo khoa học: Identification of critical active-site residues in angiotensin-converting enzyme-2 (ACE2) by site-directed mutagenesis docx
... Arg522 (NH1) coordinates the chloride ion in tACE]. Arg514 in ACE2 is displaced relative to Arg522 in ACE and is somewhat closer to the zinc-binding site. The CL2 binding site is in close proximity ... contains the zinc-bind- ing site, which faces into the deep cleft formed by the two subdomains connected at the base of the cleft. The hinge-bending motion observed upon inhibitor b...
Ngày tải lên: 20/02/2014, 01:20
Báo cáo khoa học: Investigation of the substrate specificity of a b-glycosidase from Spodoptera frugiperda using site-directed mutagenesis and bioenergetics analysis pdf
... purchased from Sigma or Merck. Site- directed mutagenesis Site- directed mutagenesis was performed using a plasmid pT7-7 [13] containing an inserted DNA fragment co ding for the mature Sfbgly50 (pT7b50) ... (dotted lines) involving the residue 39 and E451 were represented. 4-OH is represented in the equatorial (4e; pointing towards left) and axial (4a; pointing upwards) position...
Ngày tải lên: 16/03/2014, 18:20
Báo cáo Y học: Role of tyrosine 238 in the active site of Rhodotorula gracilis D-amino acid oxidase A site-directed mutagenesis study docx
... the overall substrate binding pocket remains intact. In agreement with data from the pH dependence of ligand binding and with the protein crystal structure, site- directed mutagenesis results emphasize the ... out a main role of the side chain of this active site residue in substrate/ligand fixation. The ligand-binding experiments demonstrate that the overall substrate-binding pocke...
Ngày tải lên: 17/03/2014, 17:20
Báo cáo Y học: Potential active-site residues in polyneuridine aldehyde esterase, a central enzyme of indole alkaloid biosynthesis, by modelling and site-directed mutagenesis ppt
... residue Inhibition (%) HgCl 2 (0.2) Cysteine 100 E-64 (0.025) Cysteine 0 AEBSF (1.0, 4.0) Serine 0 PMSF (1.0) Serine-Cysteine 20 TLCK (0.12) Serine-Cysteine 12 TPCK (0.2) Serine-Cysteine 0 DEPC ... primer walking using the dideoxy chain termination method [13]. For protein purification, the mutated and the wild-type cDNAs were transformed into the E. coli strain M15[pREP4]. An inoculum of .....
Ngày tải lên: 24/03/2014, 04:21
Báo cáo khoa học: Probing the determinants of coenzyme specificity in Peptostreptococcus asaccharolyticus glutamate dehydrogenase by site-directed mutagenesis potx
... nucleotide-binding protein. Nature 250, 194–199. 5 Wierenga RK, Terpstra P & Hol WG (1986) Prediction of the occurrence of the ADP-binding bab-fold in pro- teins, using an amino acid sequence ... two enzymes distinguish the cofactors in different ways [3]. In the present study, we have investigated, by means of site- directed mutagenesis and steady-state kinetics, the individ...
Ngày tải lên: 30/03/2014, 03:20
Tài liệu Báo cáo khoa học: nsights into the reaction mechanism of glycosyl hydrolase family 49 Site-directed mutagenesis and substrate preference of isopullulanase doc
... located in the vicinity of the active site and could form a binding site for a glucosyl unit i n subsite )1 (Fig. 4B). Together with the observations from the site- directed mutagenesis, Trp402 ... solution in the Dali result. It is likely that the inte rface between N-terminal and b-he lical domains participate in binding of the polysaccharide, pullulan. Comparison of the activ...
Ngày tải lên: 19/02/2014, 16:20
Tài liệu Báo cáo khoa học: Insights into the interaction of human arginase II with substrate and manganese ions by site-directed mutagenesis and kinetic studies Alteration of substrate specificity by replacement of Asn149 with Asp docx
... known to be involved in binding and hydrolysis of the guanidino group of l-arginine by arginase are strictly conserved in the act- ive site of the agmatinases [19]. Moreover, modeling studies ... agmatine in E. coli agmatinase and l-arginine in B. caldovelox arginase [37], indicating that the substrate specificity of these enzymes rely mainly in substituents at C-a. This has been,...
Ngày tải lên: 20/02/2014, 02:21