The human norepinephrine transporter(NET) gene was cloned and structurally analyzed. The far 5’ fragment containing exon 1 (a non-coding exon) and exon 2 was sequenced. The transcription start site of the gene in hum...The human norepinephrine transporter(NET) gene was cloned and structurally analyzed. The far 5’ fragment containing exon 1 (a non-coding exon) and exon 2 was sequenced. The transcription start site of the gene in human brain stem tissue was determined by primer extension analysis. It was found that the gene could be transcribed from multiple starting points. The 5’ flanking sequence contains a proximal G-C rich region, one possible GSG elemeflt and several SP1 sites. However it does not contain TATA box and CAAT box motifS. Gel shift analysis with nuclear extracts from different tissues of mouse shows that the G-C rich region may be involved in tissue specific expression of the gene.展开更多
A cDNA molecule encoding a major part of the hu-man Norepinephrine transporter(hNET) was synthesized by means of Polymerase Chain Reaction(PCR) technique and used as a probe for selecting the human genomic NET gene. A...A cDNA molecule encoding a major part of the hu-man Norepinephrine transporter(hNET) was synthesized by means of Polymerase Chain Reaction(PCR) technique and used as a probe for selecting the human genomic NET gene. A positive clone harbouring the whole gene was ob-tained from a human lymphocyte genomic library through utilizing the "genomic walking" technique. The clone, des-ignated as phNET, harbours a DNA fragment of about 59 kb in length inserted into BamH Ⅰ site in cosmid pWE15.The genomic clone contains 14 exons encoding all amino acid residues in the protein. A single exon encodes a dis-tinct transmembrane domaill, except for transmembrane domain 10 and 11, which are encoded by part of two ex-ons respectively, and exon 12, which encodes part of do-main 11 and all of domain 12. These results imply that there is a close relationship between exon splicing of a gene and structural domains of the protein, as is the case for the human γ-aminobutyric acid transporter(hGAT) and a number of other membrane proteins.展开更多
文摘The human norepinephrine transporter(NET) gene was cloned and structurally analyzed. The far 5’ fragment containing exon 1 (a non-coding exon) and exon 2 was sequenced. The transcription start site of the gene in human brain stem tissue was determined by primer extension analysis. It was found that the gene could be transcribed from multiple starting points. The 5’ flanking sequence contains a proximal G-C rich region, one possible GSG elemeflt and several SP1 sites. However it does not contain TATA box and CAAT box motifS. Gel shift analysis with nuclear extracts from different tissues of mouse shows that the G-C rich region may be involved in tissue specific expression of the gene.
文摘A cDNA molecule encoding a major part of the hu-man Norepinephrine transporter(hNET) was synthesized by means of Polymerase Chain Reaction(PCR) technique and used as a probe for selecting the human genomic NET gene. A positive clone harbouring the whole gene was ob-tained from a human lymphocyte genomic library through utilizing the "genomic walking" technique. The clone, des-ignated as phNET, harbours a DNA fragment of about 59 kb in length inserted into BamH Ⅰ site in cosmid pWE15.The genomic clone contains 14 exons encoding all amino acid residues in the protein. A single exon encodes a dis-tinct transmembrane domaill, except for transmembrane domain 10 and 11, which are encoded by part of two ex-ons respectively, and exon 12, which encodes part of do-main 11 and all of domain 12. These results imply that there is a close relationship between exon splicing of a gene and structural domains of the protein, as is the case for the human γ-aminobutyric acid transporter(hGAT) and a number of other membrane proteins.
