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首页> 外文期刊>Journal of Zhejiang University. Science, A >Genomic structure analysis of SNC6, a progesterone-receptor associated protein gene, and cloning and characterization of its 5'-flanking region
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Genomic structure analysis of SNC6, a progesterone-receptor associated protein gene, and cloning and characterization of its 5'-flanking region

机译：SNC6的基因组结构分析，孕酮受体相关蛋白基因及其5'侧翼区域的克隆和表征

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Objective: To analyze the genomic structure of SNC6, a progesterone-receptor associated protein gene and its regulatory elements in its 5'-flanking region. Methods: Genomic sequence from GenBank database (accession number: Z98048) covering the whole SNC6 gene was used to analyze the genomic structure of SNC6 and design primers for PCR amplification of its 5'-flanking region. A 1894 bp fragment of the 5'-flanking region (?1814 to +75) was cloned by PCR using genomic DNA from a healthy donor peripheral blood lymphocyte as template. This fragment, as well as 3 shorter derivative fragments (1423 bp, 632 bp and 416 bp, which correspond to ?1344 to +75, ?552 to +75 and ?337 to +75 respectively), were subcloned into pGL2 series luciferase reporter vectors. These constructs were introduced into colorectal cancer cell line SW620 for transient expression of reporter gene and luciferase activities were measured. Results: The genomic structure analysis showed there are 12 exons for SNC6 gene, which spans 32017 bp (nt71529 to nt39513 in Z98048 sequence). All transfected SW620 cells with the above 5-flanking region-containing constructs showed luciferase activities. The highest luciferase activities were measured in transfected cells with vectors containing 1894 bp fragments, and the lowest luciferase activities were measured in transfected cells with vectors containing 416 bp fragments. Luciferase activities were higher in transfected cells with vectors containing 632 bp fragments than that in transfected cells with vectors containing 1423 bp fragments. Conclusion: The basic transcription-promoting element (promoter) for SNC6 expression resides between 0 to ?337, and two transcription-enhancing elements (enhancer) resides between ?337 to ?552 and ?1344 to ?1814, whereas one transcription-inhibiting element (silencer) exists between ?552 to ?1344.
机译：目的：分析SNC6，孕激素受体相关蛋白基因及其5'侧侧侧的调节元件的基因组结构。方法：用于覆盖整个SNC6基因的Genbank数据库（登录号：Z98048）的基因组序列分析SNC6的基因组结构和其5'侧翼区域的PCR扩增设计引物。通过来自健康供体外周血淋巴细胞作为模板的基因组DNA，PCR克隆了5'-侧翼区域（α1814至+75）的1894bp片段。该片段，以及3个越短的衍生片段（1423bp，632bp和416bp，对应于α1344至+75，Δ552至+75和α337至+75），被亚克隆到Pgl2系列荧光素酶报告中vectors。将这些构建体引入结直肠癌细胞系SW620，用于记录基因的瞬时表达，并测量荧光素酶活性。结果：基因组结构分析显示SNC6基因有12个外显子，其跨越32017bp（NT71529至Z98048序列中的NT39513）。所有转染的SW620细胞具有上述含5个侧面区域的构建体显示了荧光素酶活性。在转染的细胞中测量最高的荧光素酶活性，含有1894bp片段的载体，并且在转染的细胞中测量最低的荧光素酶活性，含有416bp片段的载体。转染细胞的荧光素酶活性较高，其含有632bp片段的载体比含有1423bp碎片的转染细胞中的转染细胞。结论：SNC6表达的基本转录促进元素（启动子）在0到α37和两个转录增强元素（增强子）之间存在于α337和α352和α1344之间，而一个转录抑制元素（消音器）存在于？552到？1344之间。

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