Cloning and nucleotide sequence analysis of human embryonic zeta-globin cDNA.

Clones of human embryonic alpha-like zeta-globin cDNA were isolated, by detection using cross-hybridization to human alpha-globin cDNA probes, from a cDNA library derived from the mRNA of the human erythroleukemia cell line K562. Nucleotide sequence analysis of these cDNA clones revealed a coding sequence that corresponds perfectly to the independently derived amino acid sequence of the human zeta-globin chain. Comparison of the nucleotide sequence of human zeta-globin cDNA with that of human alpha-globin cDNA confirmed previous estimates of very distant evolutionary divergence between the human zeta- and alpha-globin genes.

Expression of herpes simplex virus type I thymidine kinase gene in Escherichia coli.

A herpes simplex virus type I DNA fragment containing the sequence coding for thymidine kinase was fused to the very beginning of the Escherichia coli lac Z gene in the three possible reading frames. When the thymidine kinase sequence was in the orientation fit to be transcribed from the lac promoter, functional thymidine kinase was made under lac control in all three cases. Sequences data indicate that translation reinitiation occurs at the 5' end of the thymidine kinase gene after stop signals.

Hemoglobin Pitie-Salpetriere beta 34 (B16) Val replaced by Phe. A new high oxygen affinity variant associated with familial erythrocytosis.

Hemoglobin Pitie-Salpetriere was detected by routine isoelectric focusing. It moved up on isoelectric focusing between hemoglobin A and hemoglobin F, near Hb F. On cellulose acetate strips at pH 8.

The human alpha-globin gene. The protein products of the duplicated genes are identical.

In order to determine whether any heterogeneity exists in the human alpha-globin chain, i.e. whether the products of the duplicated genes are identical, we have determined the total sequence of 14 alpha-globin chains: seven of these were abnormal, while six were normal chains from the same individuals, with one additional sample which consisted of the alpha chains from a normal control.

Complete amino-acid sequences of DNA-binding proteins HU-1 and HU-2 from Escherichia coli.

The DNA-binding protein HU from Escherichia coli is a heterodimer constituted of two polypeptide chains termed HU-1 and HU-2, of 90 residues each. Their primary structures were established from structural data obtained from tryptic peptides of each monomer in addition to the structural data provided by the automated Edman degradation of the dimer and by peptides derived from cleavage of the dimer with trypsin, chymotrypsin, V8 staphylococcal protease and dilute acid. The results presented in this paper confirm the amino-terminal and carboxy-terminal sequences of the dimer HU reported previously [Laine et al.

Identification of gamma-glutamyl phosphate in the alpha 2 chains of chicken bone collagen.

Purified components of chicken bone collagen contain approximately 4 atoms of organic phosphorus per mol of collagen, located principally in the alpha 2 chains. Previous analyses have demonstrated the absence of O-phosphoserine, O-phosphothreonine, and other phosphorylated hydroxy amino acids, phosphoamidated amino acids, and phosphorylated sugars. In the present report we establish that chicken bone collagen contains gamma-glutamyl phosphate.

[Different primary structure of 2 variants of Friend virus p 30 polypeptide separated by isoelectric focusing].

Variants of p 30 (iso-p 30s), the 30,000 dalton major polypeptide of murine C-type retraviruses, have been characterized in all virus strains by isoelectric focusing. Several of these iso-p 30s have been found to coexist in a given virus strain. In the present study, two iso-p 30s, characteristic of the Friend-Rauscher subgroup, separated by preparative isoelectric focusing of p 30 in thin layers of polydextran gel, were subjected to tryptic peptide mapping and aminoacid analysis.

Variability in the amount of beta-globin mRNA in beta0 thalassemia.

Globin mRNA isolated from a number of beta0 thalassemia patients of different ethnic origins was analyzed by RNA-cDNA hybridization and, in two cases, by fingerprint analysis of 125I-labeled mRNA. Quantitation of the relative amounts of alpha- and beta-mRNA by hybridization to purified alpha-and beta-cDNA revealed that in approximately half the cases, there was less than 1% as much beta-mRNA as alpha-mRNA. In the rest of the cases, low levels of beta-like mRNA were detected in amounts 4-12% as abundant as alpha-mRNA.

Functional and physicochemical studies of hemoglobin St. Louis beta 28 (B10) Leu replaced by Gln: a variant with ferric beta heme iron.

Studies have been performed on a 20-yr-old man exhibiting methemoglobinemia and a severe hemolytic anemia involving formation of Heinz bodies. This condition was due to an abnormal Hb present in the red cells of the proband: Hb St. Louis, beta 28 (B10) replaced by Gln, whose structural characteristics have been previously reported.

Nucleotide sequences of the 3'-terminal untranslated region of messenger RNA for human beta globin chain.

In normal messenger RNA for the human beta-globin chain, nucleotide sequences have been identified which can be matched to the amino-acid sequence of the abnormally long segment of the beta-chain of hemoglobin Cranston. The finding of these sequences strengthens the hypothesis that the betaCranston chain arose by a frameshift mutation allowing the "readthrough" of the normal termination codon and translation of usually untranslated portions of the messenger RNA for the beta-globin chain. The oligonucleotides which match the amino-acid sequence of hemoglobin Cranston provide a sequence of 36 nucleotides which follows the normal beta-chain termination codon UAA.

[Hemoglobin J. Broussais alpha-2 90 Lys leads to Asn beta-2A (FG2) discovered in a Martinique family. Comparison of several analytical technics].

We report a new case of hemoglobin J. Broussais, present in its heterozygote form in a seven year old child from Martinque. The structural characteristics of this abnormal hemoglobin have been compared by three methods: 1.