Isolation and characterization of cDNA clones for human and bovine fibronectins.
نویسندگان
چکیده
A bovine fibronectin (FN) cDNA clone (pFB1) was isolated by screening a cDNA library of calf testis fibroblasts with a synthetic oligonucleotide probe. The probe was a mixture of eight 14-base-long oligonucleotides designed from the amino acid sequence Glu-Cys-Phe-Met-Pro present in the Mr 3,000 COOH-terminal fragment of bovine plasma FN [Petersen, T.E., Thøgersen, H.C., Skorstengaard, K., Vibe-Pedersen, K., Sahl, P., Sottrup-Jensen, L. & Magnusson. S. (1983) Proc. Natl. Acad. Sci. USA 80. 137-141]. pFB1 contained a 1,000 base-pair (bp) insert comprising the complete 3' noncoding sequence (690 bp) and approximately equal to 300 bp of the coding region. The clone pFB1 was used as a radioactive probe in the screening of a human cell line (Hs 578T) cDNA library. Eleven positive cDNA clones were detected, one of which, named pFH1, contained a 2,000-bp insert comprising the complete 3' noncoding region (693 bp) and approximately equal to 1,300 bp of the coding region of human FN. The sequences of the clone pFB1 insert and of the homologous region in clone pFH1 were determined. The nucleotide sequences are 90% homologous. Six amino acid changes were found, clustered in an area connecting two structural domains described in bovine plasma FN. Furthermore, the 204 COOH-terminal amino acid sequence of bovine FN was completed by overlapping two peptide fragments (MrS 3,000 and 23,000). Clone pFH1 was used in estimating the size of human fibronectin mRNA (7,900 bases) through blot hybridization analysis. Southern blot studies suggest that human FN is coded by a unique gene.
منابع مشابه
Isolation and characterization of a bovine herpesvirus 3 From cases of calf pneumonia in Iran
متن کامل
Molecular Cloning and Expression of Human Gamma Interferon (IFN-g) Full cDNA in Chinese Hamster Ovary (CHO) Cells
Background: IFN-g is mostly secreted by activated CD4+ , CD8+ T cells and NK cells. This cytokine has immunomodulatory, anti-cancer and anti-microbial effects and is important for prophylaxis, diagnosis and treatment of chronic infections and cancers. Objective: The purpose of this study was to clone the full cDNA of human IFN-g and express it in CHO cell line. Methods: Lymphocytes from a healt...
متن کاملCloning and Characterization of cbhII Gene fromTrichoderma parceramosum and Its Expressionin Pichia pastoris
The genomic and cDNA clones encoding cellobiohydrolase II (CBHII) have been isolated and sequenced from a native Iranian isolate of Trichoderma parceramosum, a high cellulolytic enzymes producer isolate. This represents the first report of cbhII gene from this organism. Comparison of genomic and cDNA sequences indicates this gene contains three short introns and also an open reading frame codin...
متن کاملIsolation and characterization of microsatellite loci in the Persian sturgeon (Acipenser persicus, Borodine, 1897) and cross-species amplification in four commercial sturgeons from the Caspian Sea
In order to have a sustainable management on Persian sturgeon as a highly commercial species in the South Caspian Sea, we need to identify its population structure and the level as well as its conservation status in their natural habitat. To develop a conservation program for this all Caspian Sea' sturgeon species it requires knowledge of its genetic diversity using reliable molecular marker t...
متن کاملIsolation and characterization of 13 new microsatellite markers in the triangle mussel (Hyriopsis cumingii)
Microsatellite enriched library of Hyriopsis cumingii was constructed according to the strong affinity between biotin and streptavidin. One hundred clones with foreign inserts were sequenced and 65 clones were screened for usable microsatellites, of which 36 were deemed unique, of sufficient length (more than 8 repeats) and possessed adequate flanking regions for primer designment. Among 36 pri...
متن کاملذخیره 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 ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Proceedings of the National Academy of Sciences of the United States of America
دوره 80 11 شماره
صفحات -
تاریخ انتشار 1983