Recombinant proteins

Pwo DNA polymerase is an enzyme that is widely used in PCR due to its thermal stability and proofreading activity (3′-5′ exonuclease activity) with high accuracy. Optimizing the expression and purification of the protein is crucial to reducing the cost of production for this important enzyme. In this study, the higher expression level of the gene encoding Pwo DNA polymerase in bacteria strain E. coli BL21(DE3) was achieved by optimizing codon usage.

11p xuanphongdacy06 18-09-2024 2 1   Download

In this study, the DNA fragment encoding for the C-terminus domain of the AopB from Aeromonas hydrophila AH-1 was cloned into pET-M expression vector and expressed in Escherichia coli BL21 (DE3) host cells. The recombinant AopB-C-terminus domain was successfully purified using immobilized nickel affinity chromatography as a soluble form. Crosslinking analysis among AopB-C-terminus molecules in solution showed that this domain existed as a mixture of tetramer, trimer, dimer, and monomer forms.

6p guitaracoustic02 08-12-2021 18 3   Download

RNA polymerase II (Pol II) is a 12 subunit protein complex from yeast to human that is required for gene expression. Gdown1 containing Pol II [Pol II (G)] is a special form of Pol II that is catalytically inactive and heavily depends on the 30-subunit Mediator complex for its activator and basal dependent function in vitro. Here we report for the first time, the identification and the generation of a 15-subunit human Mediator complex via the novel multibac baculovirus expression system that is fully responsive to Pol II (G).

9p thiencuuchu 27-11-2021 21 3   Download

Fibroblast growth factor-2 (FGF-2) is a multifunctional protein that plays an important role in the regulation of proliferation, differentiation and migration of a variety of cells. The recombinant human FGF-2 (rhFGF-2) is currently used in stem cell culture, medicine and cosmetic products. In this study, we aim to produce secreted rhFGF-2 protein from a Pichia pastoris strain containing multiple copies of the fgf-2 gene to eliminate the disadvantages of intracellular expression systems

9p larachdumlanat127 02-01-2021 13 2   Download

In order to set up a system for production of recombinant proteins using suspension cell culture of Nicotiana tabacum cv. Bright Yellow 2 (BY-2), in vitro culture and genetic transformation protocols of such cells were optimised at the laboratory of Plant Cell Biotechnology, Institute of Biotechnology.

10p tamynhan8 04-11-2020 9 1   Download

In this study, the recombinant endoglucanase was purified by his - tag affinity chromatography with differrent processes, such as using phosphate buffer with or without sodium cloride, pretreatment of samples with ammonium sulphate before supplying into affinity column, using various concentration of imidazole for washing.

9p tamynhan10 24-10-2020 17 1   Download

Nghiên cứu hiệu quả điều trị bệnh Gumboro của interferon gà tái tổ hợp (recombinant chicken interferon, rChIFN) khi sử dụng chỉ mỗi interferton alpha gà (ChIFN-α) hay có sự kết hợp với interferon gamma (ChIFN-γ) được thực hiện trên gà 3 tuần tuổi.

6p vivientiane2711 28-06-2020 34 3   Download

In this study, we selected a novel gene encoding protease inhibitor from metagenome of marine sponge QT collected in Quang Tri and expressed in the Escherichia coli (E. coli) strain BL21(DE3) for finding and mining novel protease inhibitor.

8p shiwo_ding7 05-06-2019 34 1   Download

Despite high vaccination coverage after introduction of whole cell (wP) and acellular pertussis (aP) vaccines, pertussis resurgence has been reported in many countries.

9p nutifooddau 27-01-2019 18 2   Download

High-Level Recombinant Protein Production in CHO Cells Using Lentiviral Vectors and the Cumate Gene-Switch developed  an efficient system to generate in less than 2 months, starting from the cDNA, pools of CHO cells stably expressing high-level of recombinant  proteins. It is based on lentiviral vectors (LVs) for stable transduction coupled with the cumate gene-switch for inducible and efficient gene expression.

13p thuyancn 01-06-2015 35 3   Download

Bắt đầu từ những năm đầu thập niên 1970, các thành tựu về sinh học phân tử và kỹ thuật di truyền giúp chúng ta có thể hiểu rõ bản chất phân tử của các bệnh xuất hiện trên động vật, thực vật và đặc biệt là trên người. Giáo sư Paul Berg thuộc trường đại học Stanford (Hoa Kỳ), người được nhận giải Nobel hóa học năm 1980, là người đầu tiên tạo ra DNA tái tổ hợp (recombinant DNA- rDNA) vào năm 1972...

65p tangtocmuathi 23-05-2013 176 47   Download

To establish stable cell lines that produce recombinant multisubunit proteins, it is usually necessary to cotransfect cells with several independent gene constructs. Here, we show that a stepwise fusion of individually transfected cells, results in a fused cell-line that secretes a complete multisubunit protein. Functional expression of recombinant multisubunit proteins may require a defined expression ratio between each protein subunit. The cell-fusion technology described allows a predefined expression level of each subunit.

6p system191 01-06-2013 35 4   Download

The expression of mitochondrial and hydrogenosomal ADP/ATP carriers (AACs) from plants, rat and the anaerobic chytridiomycete fungus Neocallimastix spec. L2 in Escherichia coli allows a functional integration of the recombinant proteins into the bacterial cytoplasmic membrane. For AAC1 and AAC2 from rat, apparent Km values of about 40 lM for ADP, and 105 lM or 140 lM, respectively, for ATP have been determined, similar to the data reported for isolated rat mitochondria.

10p system191 01-06-2013 36 3   Download

DNA transactions in eukaryotes require that proteins gain access to target sequences packaged in chromatin. Further, interactions between distinct nucleoprotein complexes are often required to generate higher-order structures. Here, we employed two prokaryotic site-specific recombination systems to investigate how chromatin packaging affects the assembly of nucleoprotein structures of different complexities at more than 30 genomic loci. The dynamic nature of chromatin permitted protein–DNA and DNA–DNA interactions for sites of at least 34 bp in length.

7p system191 01-06-2013 50 4   Download

The isoprenoid methyl farnesoate (MF) has been implicated in the regulation of crustacean development and reproduction in conjunction with eyestalk molt inhibiting hormones and ecdysteroids. Farnesoic acid O-methyltransferase (FAMeT) catalyzes the methylation of farnesoic acid (FA) to produce MF in the terminal step of MF synthesis. We have previously cloned and characterized the shrimp FAMeT. In the present study, recombinant FAMeT (rFAMeT) was produced for bioassay and antiserum generation.

9p research12 01-06-2013 41 3   Download

Cloning and over-expression of human glucose 6-phosphate dehydrogenase (Glc6P dehydrogenase) has for the first time allowed a detailed kinetic study of a preparation that is genetically homogeneous and in which all the protein molecules are of identical age. The steady-state kinetics of the recombinant enzyme, studied by fluorimetric initial-rate measurements, gave converging linear Lineweaver–Burk plots as expected for a ternary-complex mechanism.

8p research12 01-06-2013 59 4   Download

This study investigated the enzymatic function of two putative plant GPXs, GPXle1 from Lycopersicon esculentum and GPXha2 from Helianthus annuus, which show sequence identities with the mammalian phospholipid hydroperoxide glutathione peroxidase (PHGPX). Both purified recombinant proteins expressed in Escherichia coli show PHGPX activity by reducing alkyl, fatty acid and phospholipid hydroperoxides but not hydrogen peroxide in the presence of glutathione.

7p research12 01-06-2013 37 4   Download

A cytosolic thyroid-hormone-binding protein (xCTBP), predominantly responsible for the major binding activity of T3 in the cytosol of Xenopus liver, has been shown to be identical to aldehyde dehydrogenase class 1 (ALDH1) [Yamauchi, K., Nakajima, J., Hayashi, H., Horiuchi, R. & Tata, J.R. (1999) J. Biol. Chem. 274, 8460–8469]. Within this paper we surveyed which signaling, and other, compounds affect the thyroid hormone binding activity and aldehyde dehydrogenase activity of recombinant Xenopus ALDH1 (xCTBP/xALDH1) while examining the relationship between these two activities....

8p research12 01-06-2013 38 4   Download

Chromatin is a highly dynamic structure that plays an essential role in regulating all nuclear processes that utilize the DNA template including DNA repair, replication, transcription and recombination. Thus, the mechanisms by which chromatin structures are assembled and modified are questions of broad interest. This minireview will focus on two groups of proteins: (a) histone chaperones and (b) ATPdependent chromatin remodeling machines, that co-operate to assemble DNA and histone proteins into chromatin....

7p research12 01-06-2013 32 4   Download

Murine pregnancy-associated plasma protein-A (PAPP-A) cDNA encoding a 1545 amino-acid protein has been cloned. We have also identified and cloned cDNA that encodes a novel variant of PAPP-A, PAPP-Ai, carrying a 29-residue highly basic insert. The point of insertion corresponds to a junction between two exons in the human PAPP-A gene. The human intron flanked by these exons does not encode a homologous corresponding insert, which is unique to the mouse.

10p research12 01-06-2013 42 6   Download