ABSTRACT
Highly purified thyroglobulin mRNA was isolated from human nodal euthyroid goiter. Full length cDNA was synthesized from 33S RNA by using reverse transcriptase in the presence of human placenta ribonuclease inhibitor. DNA complementary to human Tg mRNA was used in liquid hybridization experiments to determine the quantity of Tg mRNA. The amount of Tg mRNA in euthyroid nodal and congenital goiter was reduced. Tg specific mRNA was absent in thyroid cancer cells.
Subject(s)
Gene Expression , Thyroglobulin/genetics , Thyroid Diseases/genetics , Cells, Cultured , DNA/biosynthesis , DNA/genetics , Electrophoresis, Agar Gel , Goiter, Nodular/genetics , Humans , Nucleic Acid Hybridization , RNA, Messenger/genetics , RNA, Messenger/isolation & purification , Thyroid Gland/cytology , Transcription, Genetic , UltracentrifugationABSTRACT
A highly purified thyroglobulin (Tg) mRNA was isolated from human nodal euthyroid goiter. Ultracentrifugation in a 5-20% sucrose density gradient revealed that the protein has a sedimentation coefficient of 33S. cDNA was synthesized from the 33S RNA, using reverse transcriptase in the presence of a human placental ribonuclease inhibitor. In order to detect the polymorphism of restriction fragment length, screening of high molecular weight DNA from normal human thyroid gland and from nodal euthyroid and diffuse toxic goiters was carried out, using Tg cDNA probes and plasmid DNA containing a Tg cDNA fragment. After restriction with endonuclease Hind III, three Tg-containing fragments were identified both in the normal and nodal euthyroid goiters, whereas restriction with endonuclease Bam HI revealed two Tg-containing fragments in nodal euthyroid and diffuse toxic goiters.
Subject(s)
DNA/biosynthesis , RNA, Messenger/genetics , Thyroglobulin/genetics , Thyroid Diseases/genetics , Thyroid Gland/metabolism , Animals , Autoradiography , DNA Restriction Enzymes , Electrophoresis, Agar Gel , Electrophoresis, Polyacrylamide Gel , Humans , Mice , Molecular Weight , Nucleic Acid Hybridization , Plasmids , Poly A/genetics , Poly A/isolation & purification , Polymorphism, Restriction Fragment Length , RNA, Messenger/isolation & purification , Restriction Mapping , Thyroid Diseases/diagnosisABSTRACT
The pattern of BamHI fragments of DNA from three children suggested to suffer the isolated growth hormone deficiency type. IA was not different from normal pattern registered in blot hybridization with [32P]cDNA of the growth hormone gene. The data permits one to exclude the above mentioned disease that is characterized by the deletion of HGH-N gene. The analogous DNA restriction analysis using HindIII restriction endonuclease has shown, that neither the sick children, nor their parents carry the deletion in heterozygotic state. The study of normal polymorphism of the restriction fragments length has shown that as for as the frequency of polymorphic MspI restriction endonuclease sites A and B in the growth hormone gene cluster (0.67 and 0.75 respectively) is concerned the Russian population in Moscow is closer to Mediterranean one than to North-european.
Subject(s)
Dwarfism/diagnosis , Growth Hormone/genetics , DNA Restriction Enzymes , Dwarfism/genetics , Growth Hormone/deficiency , Humans , Multigene FamilySubject(s)
Collagen/genetics , DNA/biosynthesis , Globins/genetics , Thalassemia/genetics , Cloning, Molecular , HumansABSTRACT
Genetic and molecular studies on one case of alpha-thalassemia, found in Moscow region, have demonstrated that the cause of the disease is the deletion of 3 of 4 alpha-globin genes, presenting in the human diploid genome. The level of expression of the remained alpha-globin gene is much lower than that observed in the patients from other ethnic groups. One can suggest that in this case the deletion is spread on the regulatory zones of this gene.
