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4 edition of Cloning and protein expression of human kallikrein 12 found in the catalog.

Cloning and protein expression of human kallikrein 12

Nader Memari

Cloning and protein expression of human kallikrein 12

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Published by National Library of Canada in Ottawa .
Written in English

Edition Notes

Thesis (M.Sc.) -- University of Toronto, 2002.

SeriesCanadian theses = -- Th`eses canadiennes
The Physical Object
Pagination2 microfiches : negative.
ID Numbers
Open LibraryOL19898090M
ISBN 100612740404

General Sequence of Lecture Topics: Introduction, History, Review of basic gene expression Review of cloning procedures and vectors, libraries Synthetic DNA, PCR, DNA sequencing methods Reverse translation, codon usage Recombinant protein expression - prokaryotes Molecular diagnostics monoclonal antibodies, ELISA, molecular beacons, genotyping. Cloning using restriction enzymes. TA cloning and TOPO TA cloning. Recombination cloning systems: GATEWAY Cloning Technology (Invitrogen, Life Technologies) In-Fusion (Clontech, TaKaRa) Ligation independent cloning (LIC) Di- or multi-cistronic cloning. In order to isolate and amplify the vector DNA, an aliquot of the reaction mix is used for. Cloning and Protein expression Competent cells One Shot® is Invitrogen’s leading format. The box contains single-use 50µL aliquots of competent cells, as well as medium and a pUC19 positive control. One Shot® means: • Convenient format • No extra tube • No extra pipetting • No loss of competency due to repeated thaw-refreeze cycles.   A potential human homologue to rat MSP has been identified (human kallikrein 6, or hK6), based upon amino acid sequence identity (%) in comparison with the other human kallikreins (27, 30). hK6 has the highest expression in the central nervous system, breast, kidney, and uterus and may prove to be a useful biomarker for ovarian and breast.

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Cloning and protein expression of human kallikrein 12 by Nader Memari Download PDF EPUB FB2

There are many options available for protein expression from cloned DNA. These include cell-free extracts (in vitro expression systems), and bacterial, yeast, insect, and mammalian cell systems, each with its own advantages and ements for protein solubility, functionality, and yield are often the most important factors to consider when choosing an expression.

To appreciate the effect of cloning methods on protein expression, it is necessary to catalog briefly the sequences commonly added to genes that affect translation of the gene or which encode domains that make the resulting protein more useful or easier to purify ().Many of these motifs, along with cloning sites, are added to PCR primers and are thus incorporated into PCR Cited by: The Kallikrein 13 Recombinant Protein Antigen from Novus Biologicals is derived from E.

coli. The Kallikrein 13 Recombinant Protein Antigen has Cloning and protein expression of human kallikrein 12 book validated for the following applications: Antibody : ml Source: E. coli Species Reactivity: Human Applications: Antibody Competition.

Human kallikrein 6 (protease M/zyme/neurosin) was originally identified based on its aberrant expression in tumor cells and is considered a biomarker for ovarian cancer. Here, we describe the identification, cloning, and tissue expression of three novel transcript variants of the KLK6 gene that encode for wild-type kallikrein by: @article{osti_, title = {Identification and molecular cloning of novel transcripts of the human kallikrein-related peptidase 10 (KLK10) gene using next-generation sequencing}, author = {Adamopoulos, Panagiotis G.

and Kontos, Christos K. and Scorilas, Andreas}, abstractNote = {Tissue kallikrein and kallikrein-related peptidases (KLKs) form the largest group of serine. By using molecular cloning techniques, we identified a new human kallikrein gene, tentatively named KLK15 (for kallikrein 15 gene).

This new gene maps to chromosome 19q and is located between the KLK1 and KLK3 by: The human gene for cystic fibrosis, mentioned at the beginning of the chapter, was cloned by chromosome walking, and we shall examine its cloning in more detail in Chapter For any case of chromosome walking, there must be some type of criterion to assess each step of the walk for the gene of interest, and these criteria depend on the Author: Anthony Jf Griffiths, William M Gelbart, Jeffrey H Miller, Richard C Lewontin.

: Recombinant Human Kallikrein 2 Protein: Industrial & Scientific. Skip to main content. Try Prime EN Hello, Sign in Account & Lists Sign in Account & Lists Orders Try Prime Cart. Industrial & Scientific. Go Search Cyber Monday Deals Best. Synonyms: KLK1: Description: Kallikrein 1 (KLK1), also known as tissue kallikrein, is a member of the human tissue kallikrein family.

Kallikreins are a subgroup of serine proteases having diverse physiological functions. Growing evidence suggests that many kallikreins are implicated in carcinogenesis, and some have the potential to be used as cancer and other disease biomarkers.5/5(6).

n/a Ensembl ENSG n/a UniProt P n/a RefSeq (mRNA) NM_ n/a RefSeq (protein) NP_ n/a Location (UCSC) Chr – Mb n/a PubMed search n/a Wikidata View/Edit Human Kallikrein-1 is a protein that in humans is encoded by the KLK1 gene. KLK1 is a member of the peptidase S1 family. Kallikreins are a subgroup of serine Aliases: KLK1, KLKR, Klk6, hK1, kallikrein 1.

Cloning and Expression The general aim of cloning is to express specific proteins in vitro in order to study genes and gene manipulation effects.

The process starts with cloning a gene of interest (GOI) into a plasmid that contains the necessary elements for propagating the plasmid/GOI in bacteria, and later expressing the GOI in either prokaryotic or eukaryotic cells. Molecular cloning is a set of experimental methods in molecular biology that are used to assemble recombinant DNA molecules and to direct their replication within host organisms.

The use of the word cloning refers to the fact that the method involves the replication of one molecule to produce a population of cells with identical DNA molecules.

Molecular cloning generally uses DNA. the PCR product is cloned into a specific cloning or expression vector using one of the cloning methods described below.

the sequence of a positive clone is checked by sequence analysis. the gene of interest is sub-cloned into a variety of expression vectors (for different expression systems) using a fixed set of restriction enzymes or specific.

Human tissue kallikrein 9 (KLK9) is a newly identified member of the tissue kallikrein gene family. Recent reports have indicated that KLK9 mRNA is differentially expressed in ovarian and breast. Background: Kallikrein 1. Kallikrein 1 (KLK1), also known as tissue kallikrein, is a member of human tissue kallikrein family (1).

The best known physiological function of KLK1 is the cleavage of kininogen to release the vasoactive kinin peptide (bradykinin or lysyl-bradykinin), which regulates vasodilation, blood pressure reduction, smooth muscle relaxation and contraction. Cloning, expression and purification of rA2 protein from L.

infantum Then the absorbed protein was eluted with binding buffer containing mM imidazole. Finally, the purified protein was analyzed with SDS-PAGE [12].

SDS-PAGE and Western blotting SDS–PAGE was performed under denaturing and reducing conditions with a 12%. 1 CLONING, EXPRESSION AND PURIFICATION OF THE GENERAL STRESS PROTEIN YhbO FROM ESCHERICHIA COLI Jad Abdallah1, Renee Kern1, Abderrahim Malki, Viola Eckey, Gilbert Richarme* Stress molecules, Institut Jacques Monod, Universite Paris 7, 2.

Molecular Cloning: A Laboratory Manual (Fourth Edition)Molecular Cloning has served as the foundation of technical expertise in labs worldwide for 30 other manual has been so popular, or so influential.

Molecular Cloning, Fourth Edition, by the celebrated founding author Joe Sambrook and new co-author, the distinguished HHMI investigator Michael Green, preserves. Quantitative Expression of the Human Kallikrein Gene 9 (KLK9) in Ovarian Cancer A New Independent and Favorable Prognostic Marker November Cancer Research 61(21) Once protein is expressed, the protein function can be studied as it affects the cell signaling, morphology or other aspects.

Alternatively, the protein can be expressed in large quantities that can then be studied directly with other techniques.

We have a wide variety of products for use across the cloning & expression workflow. Kallikrein-5, Human Recombinant protein (code: ). Sterile Filtered clear sion system is Escherichia Coli. EcoR1& NdeIpUC 57 cloning vector pETb (+) expression vector Digestion of plasmid and expression vectors M 57 22 bp bp Verification of ligation of protein G insert M:marker,1,5,7,15,20 are colony picked after ligation Post ligation cells grown on agar+Amp.

t ligation cells grown on grown DNA extracted from the picked colony. Human kallikrein1(KLK1), also known as tissue kallikrein, is functionally conserved in its capacity to cleave the low molecular weight kininogen to release the vasoactive peptide, Lys-bradykinin which plays a role in regulating vasodilation, blood pressure reduction, smooth muscle relaxation and contraction, pain induction and inflammation.5/5(6).

Kallikrein 3, commonly known as prostate specific antigen (PSA), is a serine protease of the human tissue Kallikrein gene family (1). PSA is synthesized in the ductal and acinar epithelium of the prostate gland and secreted into the seminal plasma in high concentrations ( - 2 g/L) (2).

human protein atlas information i The Tissue Atlas contains information regarding the expression profiles of human genes both on the mRNA and protein level. The protein expression data from 44 normal human tissue types is derived from antibody-based protein profiling using immunohistochemistry.

Human kallikrein 13 (hK13; encoded by the KLK13 gene) is a secreted serine protease expressed in endocrine tissues, including the prostate, testis, breast, and ovary. We have previously reported steroid hormone regulation of the KLK13 gene and its clinical value as a marker of favorable prognosis in breast cancer at the mRNA level.

We hypothesized that hK13 Cited by: Citations for Recombinant Human Kallikrein 7 Protein, CF R&D Systems personnel manually curate a database that contains references using R&D Systems products.

The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.5/5(3). Expression and purification of recombinant Angptl2-GST. Angiopoietin-like protein 2 is a glycoprotein that is expressed in many tissues (Kim et al.

).Bacterially expressed Angptl2 is unable to stimulation expansion of hematopoietic stem cells, indicating that appropriate posttranslational modification is required for Angptl2 to be functional (Zhang et al.

).Cited by: 2. Our Kallikrein 2 Peptides and Kallikrein 2 Proteins can be used in a variety of model species: Human. Use the list below to choose the Kallikrein 2 Peptide and Kallikrein 2 Protein which is most appropriate for your research; you can click on each one to view full technical details, images, references, reviews and related products.

Protein expression using the K. lactis Protein Expression Kit - Cloning a PCR fragment into pKLAC2 (E). Introduction. The procedure below is for cloning a fragment produced by PCR into example assumes that the PCR fragment contains a 5' XhoI site, and has a stop codon, followed by a NotI site incorporated into its 3' end.

The venom fibrinogenolytic serine proteases as well as the venom plasminogen activator, share extensive sequence homology with the thrombin-like venom serine proteases such as ancrod [20, 28], batroxobin [21, 29], crotalase [30,31], gyroxin-like serine proteases, kallikrein-like enzyme from Crotalus atrox and the protein C activator from A.

c Cited by: 2. Kallikrein 6 cDNA clones are full length sequence confirmed and expression validated. There are 13 kinds of tags for each Kallikrein 6 of different species, especially GFP tag, OFP tag, FLAG tag and so on. There are three kinds of vectors for choice, cloning vector, expression vector and lentivrial expression vector.

A potential human homologue to rat MSP has been identified (human kallikrein 6, or hK6), based upon amino acid sequence identity (%) in comparison to the other human kallikreins (27,30).

hK6 has the highest expression in the central nervous system, breast, kidney, and uterus, and may prove to be. Cloning and bacterial expression of hemagglutinin antigen stem of H9N2 avian influenza virus Mohammad Amin Behzadi1,2*, Arsalan Hosseini3 and Habibollah Dadras4 1Professor Alborzi Clinical Microbiology Research Center, Shiraz University of.

Among the known human and animal tissue kallikreins, only one enzyme has the ability to release efficiently a bioactive kinin from a kininogen.

In humans, this enzyme is known as pancreatic/renal kallikrein or, with the new nomenclature, as the KLK1 gene, encoding for human kallikrein 1 (hK1 protein) (9–12).

This enzyme acts upon a liver Cited by: Toxins from Venomous Animals: Gene Cloning, Protein Expression and Biotechnological Applications. By Matheus F. Fernandes-Pedrosa, Juliana Félix-Silva and Yamara A.

Menezes. Submitted: April 2nd Reviewed: August 14th Published: July Cited by: 7. Mining human cancer datasets for kallikrein expression in cancer: the ‘KLK-CANMAP’ Shiny web tool Chenwei Wang 1, 2, Leire Moya 1, 2, Judith A.

Clements 1, 2, Colleen C. Nelson 1, 2 and Jyotsna Batra @ 1, 2Author: Chenwei Wang. DIFFICULT CLONING, PROTEIN EXPRESSION, AND ROUTINE CLONING Clone Large, Unstable, and Toxic Genes Our Difficult Cloning Competent Cell Pack contains three different competent cell types to test with difficult-to-clone DNA constructs.

Transform large plasmids using our XLGold® Kanr Ultracompetent Cellsa, which maintain the novel. Fig. Alignment of the deduced amino acid sequence of the human kallikrein pseudogene-1 (WKLK1) with members of the kallikrein multigene family.

For kallikrein protein accession numbers, please see our recent review [5]. Dashes represent gaps to bring the sequences to better alignment. The residues of theCited by: Molecular Cloning: Cloning Vectors 2. Bacteriophages: example is lambda –1/3 of genome ( Kb) is non essential –DNA is packaged into phage particles –Can only fit 40 –53 Kb of DNA –Have an in vitro packaging system –Highly efficient.

Human kallikrein 8 (hK8), whose gene was originally cloned as the human ortholog of a mouse brain protease, is known to be associated with diseases such as ovarian cancer and Alzheimer’s disease.

Recombinant human pro-kallikrein 8 was activated with lysyl endopeptidase-conjugated beads. Amino-terminal sequencing of theCited by:   The yield of the over-expressed protein after purification was mg/L (cell culture). The human CD98 cDNA amplified from Imagene plasmid was cloned in pGEX-4T1.

The construct pGEX-4T1-hCD98 was used to express the GST-hCD98 protein in the Rosetta(DE3)pLysS strain of E.

coli. The highest level of expression was detected in this case Cited by: protein expression summary i For each cancer, color-coded bars indicate the percentage of patients (maximum 12 patients) with high and medium protein expression level.

The cancer types are color-coded according to which type of normal organ the cancer originates from.