本实验室前期研究表明,小菜蛾通过有丝分裂原激活的蛋白激酶(mitogen-activated protein kinase,MAPK)信号途径上游转录激活的关键基因MAP4K4反式调控多个中肠受体基因的表达,从而介导其对苏云金芽胞杆菌Bacillus thuringiensis(Bt)产生...本实验室前期研究表明,小菜蛾通过有丝分裂原激活的蛋白激酶(mitogen-activated protein kinase,MAPK)信号途径上游转录激活的关键基因MAP4K4反式调控多个中肠受体基因的表达,从而介导其对苏云金芽胞杆菌Bacillus thuringiensis(Bt)产生的Cry1Ac杀虫蛋白的高抗性。为进一步明确MAP4K4基因转录激活而过量表达的顺式调控机制,本文利用小菜蛾基因组数据库中MAP4K4基因序列信息,首先克隆了Bt Cry1Ac敏感小菜蛾种群MAP4K4基因上游5′-侧翼序列,得到了两种不同形式的5′-侧翼序列:MAP4K4-1和MAP4K4-2。随后,预测分析了其中的潜在功能顺式作用元件,同时发现其中核苷酸的转换会导致顺式作用元件的改变。本研究初步揭示了小菜蛾MAP4K4基因的5′-侧翼序列的遗传多样性,为后续明确MAP4K4的顺式调控机制奠定了基础。展开更多
The NS 3 5′ terminal gene was amplified by reverse transcription polymerase reaction (RT PCR), and half nest polymerase reaction. The amplified fragments were cloned and sequenced. Comparing and analysing the nucleot...The NS 3 5′ terminal gene was amplified by reverse transcription polymerase reaction (RT PCR), and half nest polymerase reaction. The amplified fragments were cloned and sequenced. Comparing and analysing the nucleotide sequences and deduced amino acid sequences of the HCLV isolated from blood and other HCV strains were performed on the computer with Goldkey software. The results showed that the highest degree of nucleotide homology exists between HCLV isolated from blood and “C” strain (sk 6 cellular virus). The homology of amino acid sequence was all at or over 98% between each pair strains. It reveals that HCLV has very close relationship with “C” strain, and also proves that NS 3 gene is highly conservative in pestiviruses.展开更多
OBJECTIVE: To screen the 5' regulatory region of the aldose reductase (AR) gene for genetic variabilities causing changes in protein expression and affecting the promoter function. METHODS: The screenings were car...OBJECTIVE: To screen the 5' regulatory region of the aldose reductase (AR) gene for genetic variabilities causing changes in protein expression and affecting the promoter function. METHODS: The screenings were carried out by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP). All SSCP variants were submitted for DNA sequencing and inserted into the plasmid chloromycetin acetyl transferase (CAT) enhancer vector. The constructs were used to transfect Hela cells, and CAT assays were performed to assess promoter activity. Gel mobility shift and footprinting assays were also performed to determine the interaction between the DNA and nuclear proteins. RESULTS: Two polymorphisms, C (-106) T and C (-12) G, were identified in the regulatory region in 123 Chinese control subjects and 145 patients with type 2 diabetes mellitus. The frequencies of genotypes WT/WT, WT/C (-12) G and WT/C (-106) T were not significantly different between the subjects and patients. In the patients with and without retinopathy, frequencies of WT/C (-106) T were 31.5% and 17.5% (P 0.05) respectively. The total frequency of WT/C (-12) G and WT/C (-106) T in patients with retinopathy was 41.8%, significantly higher than that (20.0%) in patients without retinopathy (P展开更多
文摘本实验室前期研究表明,小菜蛾通过有丝分裂原激活的蛋白激酶(mitogen-activated protein kinase,MAPK)信号途径上游转录激活的关键基因MAP4K4反式调控多个中肠受体基因的表达,从而介导其对苏云金芽胞杆菌Bacillus thuringiensis(Bt)产生的Cry1Ac杀虫蛋白的高抗性。为进一步明确MAP4K4基因转录激活而过量表达的顺式调控机制,本文利用小菜蛾基因组数据库中MAP4K4基因序列信息,首先克隆了Bt Cry1Ac敏感小菜蛾种群MAP4K4基因上游5′-侧翼序列,得到了两种不同形式的5′-侧翼序列:MAP4K4-1和MAP4K4-2。随后,预测分析了其中的潜在功能顺式作用元件,同时发现其中核苷酸的转换会导致顺式作用元件的改变。本研究初步揭示了小菜蛾MAP4K4基因的5′-侧翼序列的遗传多样性,为后续明确MAP4K4的顺式调控机制奠定了基础。
文摘The NS 3 5′ terminal gene was amplified by reverse transcription polymerase reaction (RT PCR), and half nest polymerase reaction. The amplified fragments were cloned and sequenced. Comparing and analysing the nucleotide sequences and deduced amino acid sequences of the HCLV isolated from blood and other HCV strains were performed on the computer with Goldkey software. The results showed that the highest degree of nucleotide homology exists between HCLV isolated from blood and “C” strain (sk 6 cellular virus). The homology of amino acid sequence was all at or over 98% between each pair strains. It reveals that HCLV has very close relationship with “C” strain, and also proves that NS 3 gene is highly conservative in pestiviruses.
文摘OBJECTIVE: To screen the 5' regulatory region of the aldose reductase (AR) gene for genetic variabilities causing changes in protein expression and affecting the promoter function. METHODS: The screenings were carried out by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP). All SSCP variants were submitted for DNA sequencing and inserted into the plasmid chloromycetin acetyl transferase (CAT) enhancer vector. The constructs were used to transfect Hela cells, and CAT assays were performed to assess promoter activity. Gel mobility shift and footprinting assays were also performed to determine the interaction between the DNA and nuclear proteins. RESULTS: Two polymorphisms, C (-106) T and C (-12) G, were identified in the regulatory region in 123 Chinese control subjects and 145 patients with type 2 diabetes mellitus. The frequencies of genotypes WT/WT, WT/C (-12) G and WT/C (-106) T were not significantly different between the subjects and patients. In the patients with and without retinopathy, frequencies of WT/C (-106) T were 31.5% and 17.5% (P 0.05) respectively. The total frequency of WT/C (-12) G and WT/C (-106) T in patients with retinopathy was 41.8%, significantly higher than that (20.0%) in patients without retinopathy (P
