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Cloning of a Complementary Deoxyribonucleic Acid Coding for Human Thyroxine-Binding Globulin (TBG): Existence of Two TBG Messenger Ribonucleic Acid Species Possessing Different 3′-Untranslated Regions
Author(s) -
Fukushi Kambe,
Hisao Seo,
Yoshiharu Murata,
Nobuo Matsui
Publication year - 1988
Publication title -
molecular endocrinology
Language(s) - English
Resource type - Journals
eISSN - 1944-9917
pISSN - 0888-8809
DOI - 10.1210/mend-2-2-181
Subject(s) - biology , complementary dna , microbiology and biotechnology , untranslated region , polyadenylation , cdna library , coding region , three prime untranslated region , messenger rna , nucleic acid sequence , gene , genetics
An adult human liver cDNA library constructed in expression vector, bacteriophage lambda gt11, was screened with polyclonal antibody directed against human T4-binding globulin (TBG). TBG cDNA cloned in the present study was 944 nucleotides in length. It contained approximately 70% of the coding region and complete 3'-untranslated region. When the sequence was compared with that of TBG cDNA recently cloned by I. L. Flint, T. J. Bailey, T. A., Gustafson, B. E. Markham, and E. Morkin, the 3'-untranslated region of our cDNA was 231 nucleotides shorter than their cDNA. These results indicated that two TBG mRNAs with different length of 3'-untranslated regions may exist in human liver. Indeed, Northern blot analysis revealed that two TBG mRNAs differing in the length approximately 200 base pairs were present in normal human liver as well as in human hepatoma cell line (HepG2). It was demonstrated that this size difference was due to the length of 3'-untranslated region by hybridization with a probe specific to the longer 3'-end. Together with the sequence data, it was suggested that these two TBG mRNA species may be produced by alternative processing and polyadenylation at two different sites.

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