萤火虫荧光素酶基因 c DNA真核表达载体 p GL2 可在原核细胞中高效表达 .该载体中含有 SV4 0早期启动子及荧光素酶基因 c DNA 5’端总长共 75 8bp的 DNA碱基序列 ,经计算机分析发现 ,在 SV4 0早期启动子中含有 SV4 0基因组 Hind B片断...萤火虫荧光素酶基因 c DNA真核表达载体 p GL2 可在原核细胞中高效表达 .该载体中含有 SV4 0早期启动子及荧光素酶基因 c DNA 5’端总长共 75 8bp的 DNA碱基序列 ,经计算机分析发现 ,在 SV4 0早期启动子中含有 SV4 0基因组 Hind B片断中一段长为 4 9bp的 DNA序列 ,且荧光素酶基因 c DNA编码区的 5’端含有一个原核细胞基因表达调控区相似序列 .将 p GL2 中的荧光素酶基因 c DNA(L uc)克隆入真核表达载体 p ED中 ,构建成 p ED- Luc.该载体经表达测定后 ,只在真核细胞中表达 ,在原核细胞中不表达 .这说明荧光素酶基因 c DNA编码区的 5’端的原核细胞基因表达调控区相似序列不具表达有活性的荧光素酶的功能 .经 DNA序列对比研究 ,认为 p GL2 含有的 SV4 0基因组 Hind B片段 5 12 3bp~ 5 171bp长 4 9bp的 DNA序列可能就是 Hind B片段的原核增强子功能的决定序列 .展开更多
Aberrant tumor suppressor gene promoter methylation was associated with the several cancers, including breast cancer, which was the common female deaths in most countries involved in Vietnam. The methylation in tumor ...Aberrant tumor suppressor gene promoter methylation was associated with the several cancers, including breast cancer, which was the common female deaths in most countries involved in Vietnam. The methylation in tumor suppressor genes, including RASSFIA, were the key targets of establishing the potential biomarkers for prognosis and early diagnosis of breast cancer. In present study, with the aim towards using the hypermethylation at CpG islands of promoter of RASSFIA as the biomarker for breast cancer in Vietnamese population, MSP (methyl specific PCR) was carried out to analyze the hypermethylation status ofRASSFIA gene in 115 samples including 95 breast cancer specimens and 20 normal breast tissues from another disease (not breast cancer). All samples were obtained from Ho Chi Minh City Medical Hospital, Vietnam. The known predictive and prognostic factors: HER2/neu overexpression was immunohistochemistry stained as input value for breast cancer specimens. For input value confirmed, the overexpression of p53 protein was also analyzed together with prior immunochemical assay. The results indicated that the hypermethylation of frequencies for methylation of given gene reached 42.1% (P 〈 0.05). In addition, the DNA hypermethylation of RASSFIA gene increased the possibility to be breast cancer with high incidence via calculated of odd ratio (P 〈 0.05). In conclusion, the hypermethylation of candidate genes could be used as the promising biomarkers applying in Vietnamese breast cancer patients.展开更多
文摘萤火虫荧光素酶基因 c DNA真核表达载体 p GL2 可在原核细胞中高效表达 .该载体中含有 SV4 0早期启动子及荧光素酶基因 c DNA 5’端总长共 75 8bp的 DNA碱基序列 ,经计算机分析发现 ,在 SV4 0早期启动子中含有 SV4 0基因组 Hind B片断中一段长为 4 9bp的 DNA序列 ,且荧光素酶基因 c DNA编码区的 5’端含有一个原核细胞基因表达调控区相似序列 .将 p GL2 中的荧光素酶基因 c DNA(L uc)克隆入真核表达载体 p ED中 ,构建成 p ED- Luc.该载体经表达测定后 ,只在真核细胞中表达 ,在原核细胞中不表达 .这说明荧光素酶基因 c DNA编码区的 5’端的原核细胞基因表达调控区相似序列不具表达有活性的荧光素酶的功能 .经 DNA序列对比研究 ,认为 p GL2 含有的 SV4 0基因组 Hind B片段 5 12 3bp~ 5 171bp长 4 9bp的 DNA序列可能就是 Hind B片段的原核增强子功能的决定序列 .
文摘Aberrant tumor suppressor gene promoter methylation was associated with the several cancers, including breast cancer, which was the common female deaths in most countries involved in Vietnam. The methylation in tumor suppressor genes, including RASSFIA, were the key targets of establishing the potential biomarkers for prognosis and early diagnosis of breast cancer. In present study, with the aim towards using the hypermethylation at CpG islands of promoter of RASSFIA as the biomarker for breast cancer in Vietnamese population, MSP (methyl specific PCR) was carried out to analyze the hypermethylation status ofRASSFIA gene in 115 samples including 95 breast cancer specimens and 20 normal breast tissues from another disease (not breast cancer). All samples were obtained from Ho Chi Minh City Medical Hospital, Vietnam. The known predictive and prognostic factors: HER2/neu overexpression was immunohistochemistry stained as input value for breast cancer specimens. For input value confirmed, the overexpression of p53 protein was also analyzed together with prior immunochemical assay. The results indicated that the hypermethylation of frequencies for methylation of given gene reached 42.1% (P 〈 0.05). In addition, the DNA hypermethylation of RASSFIA gene increased the possibility to be breast cancer with high incidence via calculated of odd ratio (P 〈 0.05). In conclusion, the hypermethylation of candidate genes could be used as the promising biomarkers applying in Vietnamese breast cancer patients.