The Experts below are selected from a list of 65520 Experts worldwide ranked by ideXlab platform
Liu Fangxin - One of the best experts on this subject based on the ideXlab platform.
-
construction and sequence analysis of the transcriptional small hairpin rna Recombinant Plasmid targeting bcl 2 gene
Journal of Oncology, 2006Co-Authors: Liu FangxinAbstract:[Purpose] To study the new approach of tumor gene therapy by constructing Recombinant using Bcl-2 as targeting gene with the help of RNA interfering and by conducting sequence analysis. [Method] Two designed DNA sequences with small hairpin structure were cloned into the transcripted carrier Pgenesil-1 in order to construct Recombinant, and the Recombinant was transformed into DH5a strains.Then the sequence analysis was carried out after the Recombinant Plasmid was identified by enzyme restriction. [Results] The synthesized DNA fragment was cloned into the carrier after annealing. It was identified as an aim sequence by enzyme restriction and sequence analysis. The Recombinant pasmid targeting Bcl-2 gene was successfully constructed and the aim sequence was obtained. [Conclusion] Successfully constructing the Recombinant Plasmid provides the scientific base for searching a new gene therapy method for cancer.
Su Fang - One of the best experts on this subject based on the ideXlab platform.
-
Construction of Recombinant Plasmid affecting CUL5 in gene translation by RNA interference
Journal of Modern Oncology, 2010Co-Authors: Su FangAbstract:Objective:To construct the Recombinant Plasmid affecting CUL5 gene translation by RNA interference in nasopharyngeal carcinoma cell line CNE2.Methods:Genoma sequences of CUL5 gene was obtained from Genebank;three pairs of siRNA(small interfering RNAs) were designed with them based on the Tuschl's principle and the most effective one was selected to design and synthesize the oligonucleotide sequences.The annealed oligonucleotide fragments were inserted into the pGPU6/GFP/Neo Plasmid,and the Recombinant Plasmid was transformed into stain DH5α.The Plasmid identified by restricted incision enzymes was used for sequencing.Results:DNA sequencing indicated that the oligonucleotide fragments were correctly inserted into pGPU6/GFP/Neo Plasmid and RT-PCR showed that the Recombinant Plasmid can inhibit CUL5 gene translation.Conclusion:The vector expressing siRNA was successfully constructed.
P. Kyslík - One of the best experts on this subject based on the ideXlab platform.
-
IS2-mediated re-arrangement of the promoter sequence suppresses metabolic burden of the Recombinant Plasmid
Folia Microbiologica, 2005Co-Authors: R. Valešová, Václav Štěpánek, B. Večerek, P. KyslíkAbstract:Recombinant Plasmid pKA18 of the high expression bacterial system for penicillin amidase (‘penicillin G acylase’) bears the 3′ end region of IS 2 element. The IS 2 sequence replaces the −35 region of promoter of pga and extends up to TAGTAT box at position −10 of the promoter region. It therefore forms a hybrid promoter of pga p _pgaHT. A natural promoter p _pgaWT was not detected on any Recombinant Plasmid isolated from Recombinant strains of Escherichia coli constitutively producing penicillin amidase. PCR fragments carrying both types of promoters were cloned into the promoter-probe vector pET2 to compare their transcriptional activity: the activity of p _pgaWT was 5× higher than that of p _pgaHT. The same nucleotide “G” localized 28 nucleotides upstream of the translation start point was identified as the respective transcription start point of both mRNAs. An attempt was made to place the pga gene cloned on a Plasmid under the control of the natural promoter: not a single clone expressing penicillin amidase was found among 150 transformants. High transcriptional activity of the natural promoter together with high pga gene dosage could result in a deleterious metabolic burden of the periplasmic enzyme.
-
IS2-mediated re-arrangement of the promoter sequence suppresses metabolic burden of the Recombinant Plasmid
Folia Microbiologica, 2005Co-Authors: R. Valešová, Václav Štěpánek, B. Večerek, P. KyslíkAbstract:Recombinant Plasmid pKA18 of the high expression bacterial system for penicillin amidase (‘penicillin G acylase’) bears the 3′ end region of IS 2 element. The IS 2 sequence replaces the −35 region of promoter of pga and extends up to TAGTAT box at position −10 of the promoter region. It therefore forms a hybrid promoter of pga p _pgaHT. A natural promoter p _pgaWT was not detected on any Recombinant Plasmid isolated from Recombinant strains of Escherichia coli constitutively producing penicillin amidase. PCR fragments carrying both types of promoters were cloned into the promoter-probe vector pET2 to compare their transcriptional activity: the activity of p _pgaWT was 5× higher than that of p _pgaHT. The same nucleotide “G” localized 28 nucleotides upstream of the translation start point was identified as the respective transcription start point of both mRNAs. An attempt was made to place the pga gene cloned on a Plasmid under the control of the natural promoter: not a single clone expressing penicillin amidase was found among 150 transformants. High transcriptional activity of the natural promoter together with high pga gene dosage could result in a deleterious metabolic burden of the periplasmic enzyme.
Li Ning - One of the best experts on this subject based on the ideXlab platform.
-
Construction and identification of L-arginine transporter CAT-2 siRNA Recombinant Plasmid
Parenteral & Enteral Nutrition, 2012Co-Authors: Li NingAbstract:Objective: To construct and identificate the Recombinant Plasmid expression vector of cationic amino acid transporter-2 siRNA,and establish a foundation to study the influence of target siRNA on the expression of CAT-2 and L-arginine/iNOS/NO pathway.Methods: According to the mRNA sequences of L-arginine transporter CAT-2,siRNA design principles and BLAST Comparison,four pairs single-strand DNA of siRNA were designed and synthesized.Annealing into double-strand DNA,the four pairs of double-strand DNA of the siRNA were inserted into siRNA expression vector pcDNATM 6.2-GW/EmGFPmiR to construct siRNA expression Plasmid with vector construction kit BLOCK-iTTM Pol II miR RNAi Expression Vector Kit with EmGFP,and transfected the Recombinant Plasmid into the competent Escherichia coli DH5.After selecting positive Escherichia coli DH5,and extracting the Recombinant Plasmids,the sequences of the Recombinant Plasmids were finally identificated.Results: The sequencing results of the Recombinant Plasmid expressing CAT-2 siRNA confirmed construction successfully.Conclusion: The Recombinant Plasmids vectors expressing L-arginine transporter CAT-2 siRNA are successfully constructed,which establish a foundation to study the target unfluence siRNA on the expression of CAT-2 and L-arginine/iNOS/NO pathway.
Liu Fang-xin - One of the best experts on this subject based on the ideXlab platform.
-
Constructing and sequence analyzing of transcriptional small hairpin RNA Recombinant Plasmid targeting bcl-2 gene
Chongqing Medicine, 2009Co-Authors: Liu Fang-xinAbstract:Objective To construct the Recombinant Plasmids expressing connective gene bcl-2 short hair-pin RNA(shRNA) by Pgenesil-1 Plasmid vector for the further searching new gene therapy method of the tumors.Methods Two DNA sequences containing small hairpin structure were designed and synthesized.Annealing and inserted into Pgenesil-1.The Recombinant Plasmid was transformed into DH5a strain.Then the Recombinant Plasmid identified by restriction enzyme was used for sequence analysis.Results The Recombinant Plasmid targetintg bcl-2 gene was constructed and the aim sequence was obtained.Conclusion Successfully constructing the Recombinant Plasmid make it possible searching new gene therapy method of tumors.