Objective To study the cloning and sequencing of mature fragment of human bone morphogenetic protein 4 gene. Methods The template DNA was obtained from the human osteosarcoma cell line U2OS. By using RT PCR method, th...Objective To study the cloning and sequencing of mature fragment of human bone morphogenetic protein 4 gene. Methods The template DNA was obtained from the human osteosarcoma cell line U2OS. By using RT PCR method, the cDNA coding for the mature fragment of BMP 4 was amplified, cloned into the vector pUC19, and sequenced by Sanger Dideoxy mediated Chain Termination method. Results The mature fragment of BMP4 cDNA was obtained by RT PCR and determined by sequencing. Through the computer search on Genebank, the analysis showed that the homology of nucleotides and amino acids between cDNA of rhBMP4 mature fragment of this study and the published sequence was 99%. Sequence analysis showed that there were two differences, one was at base 1154(201): G→C, which had no influence on the corresponding amino acids(Val). Another was at base1222(269):C→T, the mutation at the base 1222 had the change of Ala to Val. Conclusion The mature fragment of BMP4 gene has been cloned. The results will be of great significance in treatment of skeletal injuries and diseases.展开更多
The genomic DNA is the organism’s various physiological and pathological characteristics of the material base.Single nucleotide polymorphisms(single nucleotide polymorphisms,SNPs) is the genome of the most common for...The genomic DNA is the organism’s various physiological and pathological characteristics of the material base.Single nucleotide polymorphisms(single nucleotide polymorphisms,SNPs) is the genome of the most common form of genetic variation in the human genome widespread.With the completion of the Human Genome Project to stimulate,there is growing focus on research developed in this latest 3rd generation of genetic markers of practical significance.This paper describes the classification of SNPs and characteristics of the human genome SNP s research and development and the present in today’s practice,the application of design experimts to explore the direct sequencing of amplified using PCR-based method showed that SNPs.展开更多
目的通过分析糖耐量受损(IGT)大鼠肠道菌群的差异性,初步探讨化瘀温胆汤对IGT大鼠肠道菌群多样性及丰度的干预情况。方法除空白组(n=10)的SD大鼠,其余SD大鼠予高脂高糖饮食诱导IGT大鼠模型,造模成功后随机分为模型组、化瘀温胆汤组和二...目的通过分析糖耐量受损(IGT)大鼠肠道菌群的差异性,初步探讨化瘀温胆汤对IGT大鼠肠道菌群多样性及丰度的干预情况。方法除空白组(n=10)的SD大鼠,其余SD大鼠予高脂高糖饮食诱导IGT大鼠模型,造模成功后随机分为模型组、化瘀温胆汤组和二甲双胍组(n=10),化瘀温胆汤组给予化瘀温胆汤煎剂1.25 g/mL灌胃,二甲双胍组予5 mg/mL二甲双胍悬浊液灌胃,药物干预4周后采集结肠内粪便。利用QIAamp DNA Stool Mini Kit试剂盒进行粪便样本DNA的提取,分析PCR扩增基因组DNA,构建Illumina平台文库并测序,通过生物信息分析软件对测序结果进行分析,比较各组样本菌群多样性及丰度。结果Rank-Abundance曲线显示菌群均匀度和菌群丰度从高到低依次为空白组、化瘀温胆汤组、二甲双胍组、模型组;α多样性指数结果显示与空白组比较,模型组大鼠ACE、Chao1、Shannon、Simpson指数明显降低;经化瘀温胆汤组、二甲双胍组干预后上述指数明显升高;β多样性分析结果显示空白组与模型组各自相对聚集,区分良好,化瘀温胆汤、二甲双胍组虽与空白组、模型组有部分聚集,但整体可以区分开;物种相对丰度结果显示:(1)在门水平下,各样本以Firmicutes(厚壁菌门)为优势菌,Saccharibacteria及Euryarchaeota(广古菌门)在空白组数量最多,模型组数量最少;(2)在属水平下,模型组Lactobacillus(乳酸菌属)数量最多,在空白组数量最少;通过Metastats差异分析显示,在属水平上化瘀温胆汤组Blautia(布劳特氏菌属)、Streptococcus(链球菌属)的相对丰度明显低于模型组。结论IGT大鼠肠道菌群多样性及丰度均降低,化瘀温胆汤可以调节IGT大鼠肠道菌群,降低IGT模型大鼠Blautia(布劳特氏菌属)及Streptococcus(链球菌属)的丰度。展开更多
文摘Objective To study the cloning and sequencing of mature fragment of human bone morphogenetic protein 4 gene. Methods The template DNA was obtained from the human osteosarcoma cell line U2OS. By using RT PCR method, the cDNA coding for the mature fragment of BMP 4 was amplified, cloned into the vector pUC19, and sequenced by Sanger Dideoxy mediated Chain Termination method. Results The mature fragment of BMP4 cDNA was obtained by RT PCR and determined by sequencing. Through the computer search on Genebank, the analysis showed that the homology of nucleotides and amino acids between cDNA of rhBMP4 mature fragment of this study and the published sequence was 99%. Sequence analysis showed that there were two differences, one was at base 1154(201): G→C, which had no influence on the corresponding amino acids(Val). Another was at base1222(269):C→T, the mutation at the base 1222 had the change of Ala to Val. Conclusion The mature fragment of BMP4 gene has been cloned. The results will be of great significance in treatment of skeletal injuries and diseases.
文摘The genomic DNA is the organism’s various physiological and pathological characteristics of the material base.Single nucleotide polymorphisms(single nucleotide polymorphisms,SNPs) is the genome of the most common form of genetic variation in the human genome widespread.With the completion of the Human Genome Project to stimulate,there is growing focus on research developed in this latest 3rd generation of genetic markers of practical significance.This paper describes the classification of SNPs and characteristics of the human genome SNP s research and development and the present in today’s practice,the application of design experimts to explore the direct sequencing of amplified using PCR-based method showed that SNPs.
文摘目的通过分析糖耐量受损(IGT)大鼠肠道菌群的差异性,初步探讨化瘀温胆汤对IGT大鼠肠道菌群多样性及丰度的干预情况。方法除空白组(n=10)的SD大鼠,其余SD大鼠予高脂高糖饮食诱导IGT大鼠模型,造模成功后随机分为模型组、化瘀温胆汤组和二甲双胍组(n=10),化瘀温胆汤组给予化瘀温胆汤煎剂1.25 g/mL灌胃,二甲双胍组予5 mg/mL二甲双胍悬浊液灌胃,药物干预4周后采集结肠内粪便。利用QIAamp DNA Stool Mini Kit试剂盒进行粪便样本DNA的提取,分析PCR扩增基因组DNA,构建Illumina平台文库并测序,通过生物信息分析软件对测序结果进行分析,比较各组样本菌群多样性及丰度。结果Rank-Abundance曲线显示菌群均匀度和菌群丰度从高到低依次为空白组、化瘀温胆汤组、二甲双胍组、模型组;α多样性指数结果显示与空白组比较,模型组大鼠ACE、Chao1、Shannon、Simpson指数明显降低;经化瘀温胆汤组、二甲双胍组干预后上述指数明显升高;β多样性分析结果显示空白组与模型组各自相对聚集,区分良好,化瘀温胆汤、二甲双胍组虽与空白组、模型组有部分聚集,但整体可以区分开;物种相对丰度结果显示:(1)在门水平下,各样本以Firmicutes(厚壁菌门)为优势菌,Saccharibacteria及Euryarchaeota(广古菌门)在空白组数量最多,模型组数量最少;(2)在属水平下,模型组Lactobacillus(乳酸菌属)数量最多,在空白组数量最少;通过Metastats差异分析显示,在属水平上化瘀温胆汤组Blautia(布劳特氏菌属)、Streptococcus(链球菌属)的相对丰度明显低于模型组。结论IGT大鼠肠道菌群多样性及丰度均降低,化瘀温胆汤可以调节IGT大鼠肠道菌群,降低IGT模型大鼠Blautia(布劳特氏菌属)及Streptococcus(链球菌属)的丰度。
