检索结果-南通市图书馆

Cloning and expression of canine clotting factor Ⅸ cDNA in vitro mediated by retroviral vector

在线全文

学校读者我要写书评

暂无评论

Science China(Life Sciences) 1999年第4期42卷 370-375页

作者：高啸波邱信芳卢大儒薛京伦 1. Institute of Genetics Fudan University 200433 Shanghai China

Oligonucleotide of cFIX cDNA (canine FIX, cFIX) was used to transcript mRNA of dog liver cell to cD-NA by RT-PCR, and further construct it on the plasmid vector pGEM-T. The correct sequence of cFIX cDNA was obtained which covered the entire cFIX coding region. Furthermore, GINaCcIX (driven by hCMV promoter) and GlNaMBcLX (driven by MCK enhancer and β-actin promoter) were constructed using the retroviral vector backbone of GINa. Canine skin fibroblast (CSF) was used as target cell, transduced with the above constructors respectively. The results showed that these modified CSF cells could express cFIX and that the expression levels were 173 ng/106 cell/24 h (GINaCcIX) and 211 ng/106 cell/24 h (GINaMBcIX) respectively. Those data offered a promising result for further animal study.

关键词： canine clotting factor IX canine skin fibroblast retroviral vector gene transfer gene expression.

Construction and expression of inverted configuration of retroviral vector containing intron 1 of hFIX

在线全文

学校读者我要写书评

暂无评论

Chinese Science Bulletin 1998年第4期43卷 315-318页

作者： Wang Hongwei Bao Yun Xin Yongna Yang Xiaoqin Shi Qian Lu Daru Qiu Xinfang Xue Jinglun Fudan Univ Inst Genet Shanghai 200433 Peoples R China

Intron was found to play an important role in improving gene expression. To improve the human factor IX(hFIX) expression level in hemophilia B gene therapy study, the retroviral vector containing intron 1 of hFIX gene was constructed in forwarded configuration, but the intron 1 was found spliced in virus particles by RT_PCR detection. So the inverted configuration vector G1NaPAi′IX was suggested and constructed on the basis of SNMBAIXm and transfected into PA317. Then C2C12 cells were transfected using the above virus supernatant and the G418_resistant clones were selected. PCR and RT_PCR detection found that intron 1 structure existed in C2C12 clones and retroviral particles. And the expression level of inverted vector was 3 times higher than that of forwarded vector. These results showed that the inverted configuration vector was in deed able to avoid splicing of intron 1 during the process of retroviral packaging and improved the expression level of hFIX protein.

关键词： hFIX cDNA intron 1 retroviral vector gene therapy.

Transduction of primary rat hepatocytes with bicistronic retroviral vector

在线全文

学校读者我要写书评

暂无评论

World Journal of Gastroenterology 2000年第5期6卷 725-729页

作者： Qing Xie Dan Liao Xia Qiu Zhou Shu Bing Qian Shi Shu Cheng Department of Infectious Disease Ruijin HospitalShanghai Second Medical UniversityShanghai 200025China Hurnan Genetic Therapy Research Center Shanghai Second Medical UniversityShanghai 200025China

INTRODUCTIONHepatocellular transplantation (HCT) could providea therapeutic alternative to orthotopic livertransplantation(OLT) in the treatment of hepaticmetabolic defects and experimental *** appropriate conditions,... 详细信息

关键词： primary hepatocyte recombinant retroviral vector genetic markers gene transfer hepatocellular transplantation polymerase chain reaction

Construction and expression of retroviral vector pLEGFP-N1-TERT in preparation of seed cells for skin tissue engineering

维普期刊数据库评论

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

Asian Pacific Journal of Tropical Medicine 2013年第12期6卷 960-963页

作者： Ting Tan Zhi-Qi Hu Epartment of Burn And Plastic Surgery Fuzhou General Hospital of Nanjing Command.PLA Department of Plastic Surgery Nan fang Hospital of Southern Medical University

Objective:To construct the retroviral vector pLEGFP-N1-telomerase reverse transcriptase(TERT)and to investigate the expression of TERT in neonatal mouse bypodermal ***:The polymerase chain reaction(PCR)-amplified TERT gene was inserted into plasmid *** positive clone was identified by restriction enzyme digestion and sequencing,then was transfected into packaging cells to produce retrovirus *** mouse hypodermal cells were infected with the virus to generate a stable cell *** TERT mRNA expression level,telomerase activity,and enhanced green fluorescent protein(EGFP)expression level were ***:retroviral vector pLEGFP-N1-TERT was constructed successfully,and a stable cell line of neonatal mouse hypodermal cells expressing EGFP was *** blot and immunohistochemical assay showed that the expression level of TERT was significantly elevated in the neonatal mouse hypodermal ***:A high titer of retrovirus pLEGFP-N1-TERT mediates high-level expression of the exogenous TERT gene in the neonatal mouse hypodermal *** protocol has potential applications for skin tissue engineering and cell transplantation therapy.

关键词： Telomerase reverse transcriptase retroviral vector Hypodermal cells

HIGH EFFICIENT TRANSFER AND EXPRESSION OF HUMAN CLOTTING FACTOR Ⅸ cDNA IN CULTURED HUMAN PRIMARY SKIN FIBROBLASTS FROM HEMOPHILIA B PATIENT BY retroviral VECTORS

维普期刊数据库评论

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

Science China Chemistry 1992年第2期35卷 183-193页

作者：戴一凡邱信芳薛京伦刘祖洞 Institute of Genetics Fudan University Shanghai 200433 PRC

To study the possibility of somatic gene therapy for hemophilia B via gene transfer to primary factor Ⅸ-deficient skin fibroblasts, we constructed four retroviral vectors containing factor Ⅸ cDNA driven by retroviral LTR promoter, SV40 early promoter and mouse MT-I promoter, respectively. These retroviral vectors were transfected into an amphotropic packaging cell line, PA317 cells, by electroporation, and a human iibrosarcoma cell line, HT1080 cells, was used to assay the factor Ⅸ-virus titers of these four virus-producing PA317 cells, which ranged from 2×10~4 to 5×10~5 cfu/ml. The factor Ⅸ proteins produced by bulk population of four virus-producing PA317 cells were determined by ELISA. Results showed that LTR promoter directed the highest production of factor Ⅸ at the rate of 584 rig/10~6 cells/24h, while SV40 early promoter and MT promoter directed about 10 and 20 times less production of factor Ⅸ than LTR promoter. The highest expressed retroviral vector XL-Ⅸ was used to infect a line of factor Ⅸ-deficie human primary skin fibroblasts, FDⅨ cells. The factor Ⅸ secretion rate of the infected FDⅨ cells was about 549 ng/10~6 cells/24h and over 75% of secreted factor Ⅸ was biologically active. We are convinced that this factor Ⅸ-deficient human primary skin fibroblast had been cured, or genetically corrected, by retroviral-mediated gene therapy in vitro.

关键词： gene therapy retroviral vector clotting factor Ⅸ.

Human clotting factor Ⅸ (hF Ⅸ ) secretion in goat milk after direct transfer with hF Ⅸ minigene into mammary gland by using retroviral vectors

在线全文

学校读者我要写书评

暂无评论

Chinese Science Bulletin 1997年第15期42卷 1308-1313页

作者： ZHANG Kezhong, WANG Hongwei, BAO Yun, LU Daru,XUE Jinglun (J. L. Hsueh), QIU Xinfang, HUANG Shuzhen,HUANG Ying, LI Bingyan, LI Hua and ZENG Yitao1. Institute of Genetics, Fudan University, Shanghai 200433, China 2 Shanghai Children’s Hospital, Shanghai Institute of Medical Genetics, Shanghai 200040, China

THE mammary gland bioreactor system of lactating animal, established by germ-linemicroinjection and embryo transplantation, has been one of the most exciting projects in thefield of biotechnology. However, this system is costly, laborious and tim-cosuming. In 1994,Johanna et al. reported that secretion of a foreign protein, human growth hormone (hGH),in milk had been achieved after direct introduction of the cDNA into the mammary gland ofgoats by replication-defective retroviral vector. In that work, the in vivo expression of

关键词： mammary-specific expression human clotting factor Ⅸ (hF Ⅸ) retroviral vectors.

Construction and identification of a retroviral vector of bcl-2 mRNA-cleaving ribozyme gene

在线全文

学校读者我要写书评

暂无评论

Journal of Medical Colleges of PLA(China) 1997年第3期12卷 227-230页

作者：赵永同朱峰王成济

The bcl-2 mRNA-cleaving ribozyme (RZ) gene was synthesized with EcoRI and BamHI sites, each on one of the terminals. The synthesized gene was cloned to the HincII site of the plasmid pGEM-3Zf(-) and the recombinant plasmid was termed 3ZRZ (+)/(-). The gene was sequenced and found to be correct, and was used for in vitro transcription and in vitro cleaving of bcl-2 mRNA. The RZ gene was cut out from the plasmid and subcloned in a retroviral vector pDOR-neo at the BamHI and EcoRI sites, and the new recombinant retroviral vector was named pDOR-RZ.

关键词： bcl-2 RNA,catalytic retroviral vector

Construction of retroviral vectors to induce a strong expression of human interferon-β gene in human hepatoma cells

在线全文

学校读者我要写书评

暂无评论

Journal of Medical Colleges of PLA(China) 1997年第1期12卷 50-54页

作者：曹广文高军杨文国戚中田杜平孔宪涛

Establishing the hepatoma cell-specific expression of human interferon gene mediated by retroviral vectors. Methods: Human interferon-β complementary DNA (IFN-β cDNA) was inserted into polylinker site of pMNSM retroviral vector to construct recombinant retroviral vector pMNSIFNB, where the transcription of IFN-β gene was driven by SV40 early region promoter, and MNAIFNB, where the transcription of IFN-β gene was driven by SV40 early region promoter regulated by α-fetoprotein enhancer. The retroviral constructs were respectively introduced into PA317 amphotropic packaging cells by means of lipofectamine mediated gene transfer procedure. The plasmids transfection efficiency was among (4-25)ｘ103 colonies/μg DNA/106 PA317 cells. The retrovirus infection efficiency was among (4. 5-500)ｘ104 Colony Forming Units (CFU)/ml. The recombinant retroviruses were used to infect human hepatoma cells, renal cell carcinoma cells and melanoma cell lines in the presence of 4 μg/ml polybrene. Results: Dot hybridization of total RNA from the neomycin resistant colonies and interferon expression assay indicated that human α-fetoprotein enhancer induced efficient and specific transcription and expression of IFN-β gene driven by the promoter of different origin in human hepatoma cells by which α-fetoprotein was highly produced. Conclusion: Cis-active element of α-fetoprotein gene can drive IFN-β gene specifically expressed in human hepatoma cells, which presents some valuable materials for the hepatoma-specific immune gene therapy.

关键词： human interferon-β retroviral vector hepatoma tissue-specific transcription regulatory sequence gene expression gene therapy

Improvement in Socio-Economic Productivity of HIV Positive Individuals on Antiretroviral Treatment in a Private Setting in South India

在线全文

学校读者我要写书评

暂无评论

World Journal of AIDS 2019年第1期9卷 23-32页

作者： Fehmida Visnegarwala Glory Alexander Ram Babu ASHA Foundation Bangalore India

PLHIV have decreased economic productivity both due to direct and indirect causes. Data from developed countries have shown that at the societal level, high costs ART are offset by increased productivity. We hypothesized that post-ART the SES would improve regardless of the baseline SES and will be sustained over time. Our objective was to perform a comprehensive SES evaluation pre/post ART initiation using an ambispective cohort study design. We used Indian household-specific SES validated tool, with score of 76 being affluent, along with clinical, ART adherence data at median of 6 and 18 months post ART, and compared using paired t-tests. Among 140 persons started on ART, with a median follow up of 22 months, 118 had Pre-ART SES data, of these: 57% were women;median age was 38 years;67% were married;89 (78%) had heterosexual sex as HIV risk;40 (34%) had major OI and/or TB at presentation. Reported self-occupation was: skilled labourers 41 (35%);12 (10%) unskilled labourers;27 (23%) housewives;26 (22%) pro-fessionals/blue collar job;1 student, 10 unemployed. The median pre-post ART CD4 cell counts were: 187 and 454 cells/cumm (P < 0.01);median body weight pre-post ART was 54 and 57 kg (P < 0.01);97% of the participants were 100% adherent. The mean Pre-ART total SES score was 37.06 (+/-10.2);and Post-ART SES score 40.62 (+10.1 P < 0.001) and these results were sustained over time and remained significant even when only monthly income was considered. Our data show a significant impact of ART on SES in a sustained manner in a developing world setting, which has policy level implications.

关键词： Persons Living with HIV (PLHIV) Anti retroviral Treatment (ART) Socio-Economic Status (SES)