那位大侠帮我翻译以下下面的英文啊
来源:学生作业帮 编辑:作业帮 分类:英语作业 时间:2024/07/17 22:49:57
那位大侠帮我翻译以下下面的英文啊
A Klotho promoter fragment spanning nt -1830 to +7 was synthesized from human genomic DNA (Promega) by PCR using the primers 5′-gcggtaccCCAACAGACACTCCAATC-3′ (forward primer) and 5′-cggagctcGACGCGCGGTCGCTC-3′(reverse primer). KpnI and XhoI restriction sites are indicated by lowercase letters. The amplified PCR products were ligated into the KpnI and XhoI sites of the pGL3-basic vector (Promega), yielding pKlotho-Luc(-1830/+7). A series of deletion constructs of human Klotho promoter fragments were synthesized by PCR using the pKlotho-Luc(-1830/+7) plasmid as a template. Forward primer sequences were 5′-gcggtaccAGATCCTT- CACTTGTCTG-3′ (-1330 to +7), 5′-gcggtaccCCAGCAGACCTGTGCGCA- 3′ (-930 to +7), 5′-gcggtaccCCCGGCAGGATCCCG-3′ (-415 to +7), 5′-gcggtaccGGCGACGCCTGCCGCA-3′ (-230 to +7), 5′-gcggtaccAGC- GGGGGTGGGCGCG-3′ (-90 to +7), 5′-gcggtacc CGCGGGCATAAAGG- GGCGC-3′ (-45 to +7). KpnI restriction sites are indicated by lowercase letters. The PCR products were digested with KpnI andXhoI, and ligated into the KpnI and XhoI sites of the pGL3-basic vector. The mutant construct containing point mutations for the Egr-1-binding motif (GGCGGGGCG à GGCTTGGCG) in the pKlotho-Luc (-90/+7) was generated using two-step PCR reactions yielding pKlotho-Luc(-90/+7 mtEgr1). The primers used for first step are forward, 5′-gcggtaccAG- CGGGGGTGGGCGCG-3′ and reverse, 5′-TATGCCCGCGCCAAGCCGCGC- CCG-3′ (-90 to –36); forward, 5′-CGGGCGCGGCTT- GGCGCGGGCATA- 3′ and reverse, 5′-cggagctcGACGCGCGGTCGCTC-3′ (-59 to +7);second step: forward, gcggtaccAGCGGGGGTGGGCGCG-3′ and reverse, 5′-cggagctcGACGCGCGGTCGCTC-3′. All the resultant constructs were verified by DNA sequencing and by restriction enzyme digests.
A Klotho promoter fragment spanning nt -1830 to +7 was synthesized from human genomic DNA (Promega) by PCR using the primers 5′-gcggtaccCCAACAGACACTCCAATC-3′ (forward primer) and 5′-cggagctcGACGCGCGGTCGCTC-3′(reverse primer). KpnI and XhoI restriction sites are indicated by lowercase letters. The amplified PCR products were ligated into the KpnI and XhoI sites of the pGL3-basic vector (Promega), yielding pKlotho-Luc(-1830/+7). A series of deletion constructs of human Klotho promoter fragments were synthesized by PCR using the pKlotho-Luc(-1830/+7) plasmid as a template. Forward primer sequences were 5′-gcggtaccAGATCCTT- CACTTGTCTG-3′ (-1330 to +7), 5′-gcggtaccCCAGCAGACCTGTGCGCA- 3′ (-930 to +7), 5′-gcggtaccCCCGGCAGGATCCCG-3′ (-415 to +7), 5′-gcggtaccGGCGACGCCTGCCGCA-3′ (-230 to +7), 5′-gcggtaccAGC- GGGGGTGGGCGCG-3′ (-90 to +7), 5′-gcggtacc CGCGGGCATAAAGG- GGCGC-3′ (-45 to +7). KpnI restriction sites are indicated by lowercase letters. The PCR products were digested with KpnI andXhoI, and ligated into the KpnI and XhoI sites of the pGL3-basic vector. The mutant construct containing point mutations for the Egr-1-binding motif (GGCGGGGCG à GGCTTGGCG) in the pKlotho-Luc (-90/+7) was generated using two-step PCR reactions yielding pKlotho-Luc(-90/+7 mtEgr1). The primers used for first step are forward, 5′-gcggtaccAG- CGGGGGTGGGCGCG-3′ and reverse, 5′-TATGCCCGCGCCAAGCCGCGC- CCG-3′ (-90 to –36); forward, 5′-CGGGCGCGGCTT- GGCGCGGGCATA- 3′ and reverse, 5′-cggagctcGACGCGCGGTCGCTC-3′ (-59 to +7);second step: forward, gcggtaccAGCGGGGGTGGGCGCG-3′ and reverse, 5′-cggagctcGACGCGCGGTCGCTC-3′. All the resultant constructs were verified by DNA sequencing and by restriction enzyme digests.
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