Sequence analysis of a KpnI family member near the 3' end of human beta-globin gene.

We determined the complete nucleotide sequence (6125 bp) of a full-length member of human KpnI family, designated T beta G41, which is located about 3 kb downstream from the beta-globin gene. Comparison of the sequence with the KpnI family sequence compiled by Singer revealed that a new 131 bp sequence is present in the T beta G41. Hybridization analyses showed that a few thousand of human KpnI family members are carrying this additional sequence. Computer search of DNA databases for T beta G41-homologous sequence showed that some T beta G41-homologous sequences were closely associated with pseudogenes. The T beta G41 sequence also showed significant sequence homology with ChBlym-1, a transferrin-like transforming gene of chicken. Furthermore, an amino acid sequence deduced from the T beta G41 nucleotide sequence revealed a relatively-high homology to those of human transferrin and lactotransferrin.     

A regulatory sequence near the 3'' end of sea urchin histone genes.

The 3' flanking sequences of all five histone genes have been sequenced in the histone DNA clone h19 of the sea urchin Psammechinus miliaris. A large (23 bp) and a small (10 bp) conserved sequence was found by sequence comparison, some 29-40 bp downstream from the termination codon. 12 bases of the larger homology block show a dyad symmetry. The available sequences of clone h22 of the same species and those of the histone clones pSp2 and pSp17 of Strongylocentrotus purpuratus, another sea urchin species, fit well into this comparison. Two types of sequences are involved in the dyad symmetry; one is H1, H3 and H4 specific, the other is H2A and H2B specific. If these conserved sequences are transcribed, a hairpin loop could form in the RNA molecules. This secondary structure might serve as a recognition signal for a regulatory protein.     

The nucleotide sequence of the human beta-globin gene

We report the complete nucleotide sequence of the human beta-globin gene. The purpose of this study is to obtain information necessary to study the evolutionary relationships between members of the human beta-like globin gene family and to provide the basis for comparing normal beta-globin genes with those obtained from the DNA of individuals with genetic defects in hemoglobin expression.     

与人体β—珠蛋白基因5‘旁侧正调控区（PCR）相结合的调控因子（简报）

Using gel mobility shift assay, three protein factors (Pa, Pb, Pc) binding to the positive control region (PCR) in the 5' flanking sequence of human beta-globin gene were detected in the nuclear extracts from mouse fetal liver at d 18 or d 19 of gestation. Competitive experiment showed that Pb and Pc could bind to GATA-1 motif, therefore could be the members of GATA family. While Pa was a new and developmental stage specific trans-acting factor, we suggested that the factor Pa was related to the activation of beta-globin gene.     

Nucleotide sequence of the 3'' exon of the human N-myc gene.

We have analyzed a 3.8 kb Eco RI fragment of genomic DNA obtained from the amplified N-myc gene of human neuroblastoma cell line BE(2)-C. This fragment contains an exon with an open reading frame encoding approximately 170 amino acids of the carboxy-terminal end of the putative N-myc protein. Comparison of the inferred amino acid sequence of this peptide with that of the 3' domain of the human c-myc protein shows that locally conserved but dispersed regions of homology exist throughout the lengths of these peptides, while hydropathy plots indicate that the physical properties implied by their primary sequences are strikingly similar. Based upon these and other considerations, it is suggested that the 3' domains of c-myc and N-myc may potentially share related functions.     

Signal sequence for generation of mRNA 3'' end in the Saccharomyces cerevisiae GAL7 gene.

We have identified a signal sequence (designated core signal) necessary to specify formation of mRNA 3' end of the GAL7 gene in Saccharomyces cerevisiae within a DNA segment 26 bp long. The sequence was located 4-5 nucleotides upstream from the 3' end, i.e. the polyadenylation site, of the GAL7 mRNA. Replacement of a DNA segment encompassing the polyadenylation site with a pBR322 DNA, leaving the core signal intact, resulted in alteration of the mRNA 3' end by several nucleotides, suggesting the existence of an additional signal (designated end signal) at or near the polyadenylation site. The normal end formation was abolished when the core signal was placed in the reverse orientation. A considerable fraction of pre-mRNA synthesized in vitro with SP6 RNA polymerase on the template of a DNA fragment containing these signals was cleaved and polyadenylated presumably at the in vitro 3' end during incubation in a cell-free system of yeast. By contrast pre-mRNA synthesized on the template with the core signal alone was processed but much less efficiently. No such processing was seen when the pre-mRNA either lacked the core signal or contained it in the reverse orientation.     

Tissue-specific expression is conferred by a sequence from the 5'' end of the rat albumin gene.

We have constructed a transient expression vector containing 400 bp of rat albumin gene immediate 5'-flanking sequences inserted 5' to the bacterial enzyme chloramphenicol acetyl transferase (CAT). We have transfected various clones of rat hepatoma cells representing different states of expression of the liver phenotype with this vector (pALB-cat) and also with two control vectors containing viral promoters (pSVE-cat and pRSV-cat), and measured activity of the bacterial enzyme CAT in cellular extracts 48 h later. The albumin flanking sequences are able to direct highly efficient CAT expression, compared with the control vectors, only in cells which express their own albumin gene: the albumin-negative hepatoma cells are at least 100 times less efficient in expressing CAT after transfection with the pALB-cat plasmid than are the albumin-positive ones. An unexpected result of our study is the total inability of the rat albumin flanking sequences to direct expression in albumin-producing mouse hepatoma cells.     

Nucleotide sequence at the 5'' end of ovalbumin messenger RNA from chicken.

DNA-sequence analysis of 300 nucleotides from the region of cloned, double-stranded ovalbumin cDNA corresponding to the 5' end of ovalbumin messenger RNA was accomplished using the technique of Maxam and Gilbert (Proc. Nat. Acad. Sci. USA (1977) 74,560-564). The AUG initiation codon was located 52 nucleotides from the AT linkers used in cloning and immediately adjacent to the amino terminal peptide of ovalbumin, indicating the absence of a "signal peptide" in this protein. The nucleotide sequence coding for a phosphorylated peptide from ovalbumin was also found. These results demonstrate that the coding portion of mRNAov begins near the 5' end of the molecule leaving some 600 nucleotides of noncoding information at the 3' end.     

The relationships between the 5'' end repeats and the largest members of the L1 interspersed repeated family in the mouse genome.

Analysis of a few large L1 elements has revealed two types of tandem repeats at the 5' end: A and F. In this study, the relationships between these repeats and a series of large L1 elements has been analysed. Most of cloned L1 repeats were shown to lack either A or F sequences at their 5' ends. F sequences are found less frequently associated than A sequences to the 5' ends of L1 and an evolutionary comparison shows that the A type was introduced more recently during the evolution of the mouse genome than the F type.     

The structure of the human beta-globin gene in beta-thalassaemia.

Twenty-one cases of beta 0 and beta +-thalassaemia have been analysed by restriction endonuclease mapping. In most cases no deletion in the regions surrounding the beta- and delta-globin genes could be detected. However, in a single Asian case of beta 0-thalassaemia, homozygous clinically, one of the homologous chromosomes contained a beta-globin gene with a deletion of 600 base pairs of DNA and comprising most or all of the 3' end of the structural gene including the EcoRI restriction site within the beta-globin coding sequence.     

Nucleotide sequence of the 3' nuclease-sensitive region of the human phosphoglycerate kinase 1 (PGK1) gene.

Many genes are known to have nuclease-sensitive sites and/or control sequences in their 3' flanking regions, but for very few genes has this region been sequenced. Previously, we mapped specific, gene activity-dependent DNAase I- and MspI-sensitive sites at the 3' end of the human X-linked housekeeping gene phosphoglycerate kinase (PGK1). Sequence information presented here shows that the 3' nuclease-sensitive site maps precisely to an Alu sequence and near a "BKM" repeat. This is the first report of an Alu sequence that has alternative chromatin configurations depending on gene activity.