Gene cloning

Bacillus velezensis RB.IBE29 was originally isolated from the rhizospheric soil of black pepper cultivated in the Central Highlands. Previous studies showed that this bacterium was a good chitinase producer, biocontrol agent, and biofertilizer.

6p viengfa 28-10-2024 3 0   Download

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

Glutamine synthetase (EC 6.3.1.2) is an enzyme which assimilates ammonium into amino acids. Therefore, GS plays an essential role in the metabolism of nitrogen. In vascular plants, GS is devided into two groups: cytosol - located isoform (GS1) and plastid – located isoform (GS2). Surveys have shown that GS1 gene was used widely to generate transgenic plants. Effects on the growth characteristics of GS1 transgenic plants have been reported in many researches.

7p vilarry 01-04-2024 8 2   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

The purpose of the study was to measure the secretion levels of recombinant glucoamylase from Pichia pastoris, by using the α-mating factor secretion signal peptide (α-MF) and the native signal peptide of glucoamylase from Aspergillus flavus NSH9. The Aspergillus flavus NSH9 gene (with and without native signal sequences), encoding a pH and thermostable glucoamylase with an SBD, was successfully cloned and expressed in Pichia pastoris to produce recombinant glucoamylases.

4p mudbound 10-12-2021 11 1   Download

In this study, the Mgra16281 gene (ID: MK322955.1) enco ding an effector with the unknown function was cloned from the rice ro ot-knot nematode Meloidogyne graminicola isolated in Long An province. To knock-down the expression of this gene, an artificial microRNA was synthesized based on the Osa-MIR528 precursor and inserted into an expression vector.

9p larachdumlanat129 19-01-2021 15 2   Download

Coffee plant is one of the most important industrial crops, and the two popular cultivars, Coffea arabica and Coffea canephora, contribute to the production of almost all coffee beans around the world. Traditional breeding could be used to develop new coffee cultivars with a higher productivity under these harsh conditions, and a biotechnological approach can also be used to improve coffee plants in a relatively short period of time.

11p baoanhlib 24-01-2017 83 3   Download

Chapter 4 review the fundamentals of gene structure and function, we are ready to start a more detailed study of molecular biology. The main focus of chapter will be the experiments that molecular biologists have performed to elucidate the structure and function of genes.

67p tangtuy05 30-03-2016 93 6   Download

Chapter 5 - Molecular tools for studying genes and gene activity. This chapter describe the most popular techniques that molecular biologists use to investigate the structure and function of genes. Most of these start with cloned genes. Many use gel electrophoresis. Many also use labeled tracers, and many rely on nucleic acid hybridization. We have already examined gene cloning techniques.

82p tangtuy05 30-03-2016 37 2   Download

Bài giảng "Simple cloning" cung cấp cho người học các kiến thức: Vector pUC, vai trò của các gene, cấu trúc của các gen, cắt vector và DNA, nối bằng ligase được, chuyển tất cả cấu trúc vào E.coli, chọn dòng mang DNA tái tổ hợp, vector nhân tạo đầu tiên, tế bào chủ, tính chất của tế bào chủ, nhân dòng sản phẩm PCR,... Mời các bạn cùng tham khảo.

55p doinhugiobay_12 15-01-2016 55 5   Download

Bài giảng "Công nghệ di truyền - Chương 3: Simple cloning" cung cấp cho người học các kiến thức: Vector pUC, vai trò của các gene, cắt vector và DNA, nối bằng ligase được, chuyển tất cả cấu trúc vào E.coli, Operon lacZ và IPTG, chọn dòng mang DNA tái tổ hợp,... Mời các bạn cùng tham khảo nội dung chi tiết.

55p doinhugiobay_10 12-01-2016 111 19   Download

An optogenetic upgrade for the tet - OFF system fused the inhibitor to the blue light-responsive B-LID degron and optimized the performance of this construct with regard to the number of Med25VBD repeats. This approach resulted in an optogenetic upgrade of the popular Tet-OFF (TetR-VP64, tetO7-PhCMVmin)  system  that  allows tunable, blue light-inducible gene expression in HEK-293T  cells.

5p thuyancn 01-06-2015 30 5   Download

The plastid transcription kinase (PTK), a component of the major RNA polymerase complex from mustard chloroplasts, has been implicated in redox-mediated regulation of plastid gene expression. A cloning strategy to define the PTK gene(s) resulted in the isolation of a full-length cDNA for a protein with overall high homology with the a subunit of cytosolic casein kinase (CK2) that contained an N-terminal extension for a putative plastid transit peptide.

9p system191 01-06-2013 45 4   Download

We report here the full coding sequence of a novel mouse putative membrane-associated mucin containing three extracellular EGF-like motifs and a mucin-like domain consisting of at least 20 tandem repeats of 124–126 amino acids. Screening a cosmid and a BAC libraries allowed to isolate several genomic clones. Genomic and cDNA sequence comparisons showed that the gene consists of 25 exons and 24 introns covering a genomic region of  52 kb. The first intron is  16 kb in length and is followed by an unusually large exon ( 9.5 kb) encoding Ser/Thr-rich tandemly repeated sequences....

10p system191 01-06-2013 44 4   Download

Manganese lipoxygenase was isolated to homogeneity from the take-all fungus, Gaeumannomyces graminis. The C-terminal amino acids and several internal peptides were sequenced, and the information was used to obtain a cDNA probe by RT/PCR. Screening of a genomic library of G. graminis yielded a full-length clone of the Mn-Lipoxygenase gene. cDNA analysis showed that the gene spanned 2.6 kb and contained one intron (133 bp). Northern blot analyses indicated two transcripts (2.7 and 3.1 kb). The deduced amino-acid sequence of the Mn-Lipoxygenase precursor (618 amino acids, 67.

8p system191 01-06-2013 36 4   Download

We previously reported that GTS1 is involved in regulating ultradian oscillations of the glycolytic pathway induced by cyanide in cell suspensions as well as oscillations of energy metabolism in aerobic continuous cultures. Here, we screened a yeast cDNA library for proteins that bind to Gts1p using the yeast two-hybrid system and cloned multiple TDH cDNAs encoding the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH).

10p research12 01-06-2013 40 3   Download

Cytochrome c is widely distributed in bacterial species, from mesophiles to thermophiles, and is one of the best-characterized redox proteins in terms of biogenesis, folding, structure, function, and evolution. Experimental molecular biology techniques (gene cloning and expression) have become applicable to cytochrome c, enabling its engineering and manipulation. Heterologous expression systems for cytochromes c in bacteria, for use in mutagenesis studies, have been established by extensive investigation of the biological process by which the functional structure is formed. ...

7p research12 01-06-2013 32 3   Download

Transglutaminase (TGase) catalyses the post-translational modification of proteins by transamidation of available glutamine residues. While several TGase genes of fish and arthropods have been cloned and appear to have similar structures to those of mammals, no homologous gene has been found in lower eukaryotes. We have cloned the acellular slime mold Physarum polycephalum TGase cDNA using RT-PCR with degenerated primers, based on the partial amino acid sequence of the purified enzyme. The cDNA contained a 2565-bp ORF encoding a 855-residue polypeptide.

10p research12 01-06-2013 48 4   Download

We report the cloning and functional charaterization of the full-length cDNA and gene encoding a Toxoplasma gondii DNA repair enzyme designated TgDRE. The gene is composed of three exons separated by two introns of 780 and 630 bp, and encodes a protein with a predicted molecular mass of 49.6 kDa. The native TgDRE protein, with a molecular mass of 60 kDa, is only detected in the virulent tachyzoite stage of T. gondii. However, the transcript is present in both asexual parasite stages, virulent tachyzoite and avirulent encysted bradyzoite. ...

9p research12 01-06-2013 33 5   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