Efficient transcription of a protein-coding gene from the RNA polymerase I promoter in transfected cells.

نویسندگان

  • I Grummt
  • J A Skinner
چکیده

The activity of the mouse ribosomal promoter was examined after fusion to the gene coding for chloramphenicol acetyltransferase (CAT) and transfection into mouse cells. Very little CAT enzyme but high levels of CAT-specific RNA correctly initiated at the ribosomal DNA start site were synthesized. The amount of specific transcripts was neither influenced by long stretches of upstream spacer sequences nor by the insertion of the Moloney murine sarcoma virus enhancer. The deletion mutant pMr delta-39, which has been shown to be fully active in vitro, exhibited a 90% decrease in template activity in vivo. A mutant in which 22 base pairs of ribosomal DNA (between positions -35 and -14) were substituted by foreign DNA sequences proved transcriptionally inactive. The fusion genes were only transcribed in mouse cells, indicating that species-specific transcription factors are involved in ribosomal promoter recognition.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Gene Expression under F8 Promoter Driving In Mouse Hepatoma Cells: A Step towards Gene Therapy of Hemophilia

Background and Objectives: Significant progress has been made in treatment of hemophilia. Ex-vivo gene therapy is going popular due to the capability of this method in using isogenic cells for genetic manipulation and reintroducing them into same host after proliferation. Most gene therapy techniques use viral vectors, which usually harbor a strong and non-specific promoter (e...

متن کامل

The Effect of Herpes Simplex Virus Virion Host Shutoff Gene- a New Suicide Gene- on Tumor Cells

Background: The herpes simplex virus (HSV) UL41 gene product, virion host shutoff (Vhs) protein, mediates the rapid degradation of both viral and cellular mRNA. This ability suggests that Vhs protein can be used as a suicide gene in cancer gene therapy applications. The recent reports have shown that the degradation of cellular mRNA during herpes simplex infection is selective. RNA containing A...

متن کامل

Highly Efficient Transfection of Dendritic Cells Derived from Esophageal Squamous Cell Carcinoma Patient: Optimization with Green Fluorescent Protein and Validation with Tumor RNA as a Tool for Immuno-genetherapy

This study was conducted to optimize a highly efficient mRNA transfection into dendritic cells (DC) derived from esophageal squamous cell carcinoma (ESCC) patients. Applying an electroporation technique, in vitro synthesized Green Fluorescent Protein (GFP) mRNA was transfected as an indicator into the DCs derived from a healthy donor. Flow cytometry revealed 84.9% transfection efficiency for DC...

متن کامل

Expression Analysis of Interferon Beta Level in HEK293T Cells Using Real-Time PCR and Protein Tests

Background: Interferons are some kind of natural cytokines which express in response to a variety of antigens including viral RNA, bacterial products, and tumor proteins. Interferon beta is used in the treatment of autoimmune diseases such as multiple sclerosis. Moreover, this drug inhibits cellular proliferation as well as angiogenesis and as a result, helps to cure cancer. In this research, i...

متن کامل

Functions of the Heterologous Intron-Derived Fragments Intra and Extra Factor IX-cDNA Coding Region on the Human Factor IX Expression in HepG2 and Hek-293T Cells

Background: Human FIX (hFIX) gene transfer into hepatocytes has provided a novel approach for treatment of hemophilia B. To obtain an improved expression of hFIX, the functional hFIX-expressing plasmids with appropriate intron-derived fragments which facilitate transcription and promote an efficient 3′-end formation of mRNAs are required.Objectives: We ai...

متن کامل

Developing Michigan Cancer Foundation 7 Cells with Stable Expression of E7 Gene of Human Papillomavirus Type 16

Background: Human papillomavirus (HPV) is responsible for the development of cervical neoplasia.  Infection with human papillomavirus type 16 (HPV-16) is a major risk factor for the development of cervical cancer. The virus encodes three oncoproteins (E5, E6 and E7), of which, the E7 oncoprotein is the major protein involved in cell immortalization and transformation o...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:
  • Proceedings of the National Academy of Sciences of the United States of America

دوره 82 3  شماره 

صفحات  -

تاریخ انتشار 1985