Recombinant DNA

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

Nội dung của tài liệu "Công nghệ tế bào động vật ứng dụng" gồm có Ứng dụng kỹ thuật recombinant dna và tạo dòng gene để điều chế các sản phẩm sinh học; mô da dùng trong dòng hóa; lập trình của di truyền ngoài nhân DNA; xác định tính đa năng của tế bào;...Mời các bạn cùng tham khảo nội dung phần 2 dưới đây.

105p dongcoxanh2510 21-10-2022 22 4   Download

DNA repair deficiencies are characteristic of cancer and homologous recombination deficiency (HRD) is the most common. HRD sensitizes tumour cells to PARP inhibitors so it is important to understand the landscape of HRD across diferent solid tumour types. Methods: Germline and somatic BRCA mu

13p vielonmusk 21-01-2022 17 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

In this study, we evaluated the reliability of genomic DNAs and cloned recombinant plasmids as standard controls for absolute real-time PCR assay. The associated standard curves were constructed and used for the quantification of Lactobacillus acidophilus probiotics.

4p mudbound 10-12-2021 32 2   Download

Lập bản đồ gene của DNA ty thể * Lập bản đồ bộ gene ty thể của nấm men Có các phương pháp khác nhau xây dựng bản đồ bộ gene ty thể: - Lập bản đồ tái tổ hợp (recombination mapping): Ở nấm men sự phân li tế bào chất và tái tổ hợp xảy ra trong quá trình mọc chồi ở cytohet lưỡng bội. Sự phân li và tái tổ hợp có thể phát hiện trực tiếp ở các dạng lưỡng bội mọc chồi hay quan sát sản phẩm giảm phân khi chồi được kích thích tạo bào tử. Ví dụ...

3p butmauvang 29-08-2013 103 12   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 177 47   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

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

Binding of lamin B receptor (LBR) to chromatin was studied by means of an in vitro assay system involving recombinant fragments of human LBR and Xenopus sperm chromatin. Glutathione-S-transferase (GST)-fused proteins including LBR fragments containing the N-terminal region (residues 1–53) and arginine-serine repeat-containing region (residues 54–89) bound to chromatin. The binding of GST-fusion proteins incorporating the N-terminal and arginine-serine repeat-containing regions to chromatin was suppressed by mild trypsinization of the chromatin and by pretreatment with a DNA solution....

11p research12 01-06-2013 29 4   Download

The RuvA, RuvB andRuvCproteins ofEscherichia coliact together to process Holliday junctions formed during recombination and DNA repair. RuvA has a well-de®ned DNA binding surface that is sculptured speci®cally to accommodate a Holliday junction and allow subsequent loading of RuvB and RuvC. A negatively charged pin pro-jecting from the centre limits binding of linear duplexDNA. The amino-acid sequences forming the pin are highly con-served.

9p research12 29-04-2013 39 3   Download

We have cloned the gene coding for theBacillus subtilis glycine oxidase (GO), a new ¯avoprotein that oxidizes gly-cine and sarcosine to the corresponding a-keto acid, ammonia and hydrogen peroxide. By inserting the DNA encoding for GO into the multiple cloning site of the expressionvectorpT7.7weproducedarecombinantplasmid (pT7-GO).ThepT7-GOencodes a fullyactive fusionprotein with six additional residues at the N-terminus of GO (MARIRA).

8p research12 29-04-2013 38 3   Download

Glofish is the first transgenic animal approved to be consumed by human in the USA  The insertion of different constructs of GFP into the fish genomes to give different green colors.The idea for recombinant DNA was first proposed by Peter Lobban, a graduate student of Prof. Dale Kaiser in the Biochemistry Department at Stanford University Medical School. The first publications describing the successful production and intracellular replication of recombinant DNA appeared in 1972 and 1973.

54p zingzing09 24-04-2013 77 5   Download

The RuvABC resolvasome ofEscherichia colicatalyses the resolution of Holliday junctions that arise during genetic recombination and DNA repair. This process involves two key steps: branch migration, catalysed by the RuvB protein that is targeted to the Holliday junction by the structure specific RuvAprotein, and resolution, which is catalysed by the RuvC endonuclease.

10p tumor12 22-04-2013 33 2   Download

We have isolated and studied the cloned sigma factors SASIG1-3 from mustard (Sinapis alba). In functional ana-lyses using both promoter and factor mutants,the three recombinant proteins all had similar basic properties but also revealed differences in promoter preference and requirements for single nucleotide positions. Directedmuta-genesis of SASIG1 identified critical residues within the conserved regions 2.4 and 4.2 necessary for binding of the )10 and)35 promoter elements,respectively.

13p tumor12 20-04-2013 33 2   Download

Sequence analysis of aPaenibacillussp. BP-23 recombinant clone coding for a previously described endoglucanase revealed the presence of an additional truncated ORF with homology to family 48 glycosyl hydrolases. The corres-ponding 3509-bp DNA fragment was isolated after gene walking and cloned inEscherichia coliXl1-Blue for expres-sion and purification. The encoded enzyme, a cellulase of 1091 amino acids with a deduced molecular mass of 118 kDa and a pI of 4.

7p fptmusic 16-04-2013 50 3   Download

Certain natural peptides and proteins of mammalian origin are able to bind and condense plasmid DNA, a prerequisite for the formation of transfec-tion-competent complexes that facilitate nonviral gene delivery. Here we have generated recombinant derivatives of the human high-mobility group (HMG) protein HMGB2 and investigated their potential as novel protein-based transfection reagents.

16p fptmusic 11-04-2013 40 1   Download

Replication protein A (RPA), a heterotrimeric protein composed of 70, 32 and 14-kDa subunits, has been shown to be essential for DNA replication, repair, recombination, and transcription. Previously, we found that, in two seed plants, rice and Arabidopsis, there are two different types of RPA70-kDa subunit. Substantial biochemical and genetic characterization of these two subunits, termed OsRPA70aand OsRPA70bor AtRPA70a and AtRPA70b, respectively, is described in this report.

12p fptmusic 11-04-2013 42 3   Download

TheSCO2299gene fromStreptomyces coelicolorencodes a single peptide consisting of 497 amino acid residues. Its N-terminal region shows high amino acid sequence similarity to RNase HI, whereas its C-terminal region bears similarity to the CobC protein, which is involved in the synthesis of cobalamin. The SCO2299gene suppressed a temperature-sensitive growth defect of anEscherichia coliRNase H-deficient strain, and the recombinant SCO2299 protein cleaved an RNA strand of RNAÆDNA hybridin vitro.

0p awards 06-04-2013 42 1   Download

RecA protein plays a pivotal role in homologous recombination inEscheri-chia coli. RecA polymerizes on single-stranded (ss) DNA forming a nucleo-protein filament. Then double-stranded (ds) DNA is bound and searched for segments homologous to the ssDNA. Finally, homologous strands are exchanged, a new DNA duplex is formed, and ssDNA is displaced. We report a quantitative analysis of RecA interactions with ss d(pN)n of var-ious structures and lengths using these oligonucleotides as inhibitors of RecA filamentation on d(pT)20....

0p awards 06-04-2013 34 3   Download