[Objective] The aim was to establish a molecular biological method for identifying coccidium species.[Method]First,the pure species of Eimeria intestinalis was isolated by using single-oocyst isolation technique.Then,...[Objective] The aim was to establish a molecular biological method for identifying coccidium species.[Method]First,the pure species of Eimeria intestinalis was isolated by using single-oocyst isolation technique.Then,according to the 18s rDNA and 5.8s rDNA sequences of Eimeria coccidia published in GenBank,a pair of specific primers was designed and synthesized to amplify the internal transcribed spacer 1（ITS-1）.Finally,the PCR products were sent for sequencing.[Result]The pure species of E.intestinalis was isolated and the result of agarose gel electrophoresis showed that the PCR product was 434 bp,and at least 27-sporulated oocysts could be detected.[Conclusion]The research will provide a basis for accurate identification of coccidium species and strains.展开更多
[Objective] The study aimed at cloning and analyzing the insulin-like growth factor-1 （IGF-1） gene from liver of Hubei white swine. [Method] The total RNA was extracted by using Trizol from the liver of Hubei white ...[Objective] The study aimed at cloning and analyzing the insulin-like growth factor-1 （IGF-1） gene from liver of Hubei white swine. [Method] The total RNA was extracted by using Trizol from the liver of Hubei white swine and used as template to amplify IGF-1 gene cDNA by RT-PCR. The cDNA product was cloned into pCRII vector, screened with blue-white colonies, digested with double enzymes and sequenced. [Result] The sequencing result indicated that the IGF-1 gene consisted of 607 nucleotides, containing 5＇-untranslated region at nucleotides 1-145, a complete ORF at nucleotides 146-538 encoding 130 amino acids, and 3＇-untranslated region at nucleotides 539-607. It shared 100% homology with the porcine IGF-1 gene reported by Muller et al. [Conclusion] The successful cloning and sequencing of the Hubei white swine IGF-1 gene confirmed that IGF-I gene was highly conserved, which provided technical basis for the use of transgenic technology for breeding of Hubei white swine.展开更多
目的分析基于活性导向分离茯苓中具有抑制人乳腺癌MDA-MB-231细胞迁移的成分并探索其分子机制。方法茯苓经醇提去除小分子成分,水提醇沉淀后得茯苓多糖(PPS),PPS经DEAE Sepharose Fast Flow阴离子交换树脂分离和Superdex-75系列凝胶纯...目的分析基于活性导向分离茯苓中具有抑制人乳腺癌MDA-MB-231细胞迁移的成分并探索其分子机制。方法茯苓经醇提去除小分子成分,水提醇沉淀后得茯苓多糖(PPS),PPS经DEAE Sepharose Fast Flow阴离子交换树脂分离和Superdex-75系列凝胶纯化后活性跟踪得PPS10-2,并对PPS10-2进行高效凝胶渗透色谱(HPGPC)法分析、单糖组成测定和活性及机制探索。结果PPS抑制人乳腺癌MDA-MB-231细胞迁移,采用活性跟踪方法,从茯苓中得到活性均一多糖PPS10-2,相对分子量为5761D,主要由鼠李糖∶阿拉伯糖∶葡萄糖组成,三种单糖的摩尔比为10.0∶3.3∶7.0,PPS和PPS10-2表现出较强的抑制MDA-MB-231细胞迁移能力,并且PPS和PPS10-2均是通过抑制特异富含AT序列结合蛋白(SATB)-1的表达来抑制MDA-MB-231细胞迁移能力,推测PPS中可能是PPS10-2为主要成分发挥该作用。结论从茯苓中成功分离到具有抑制MDA-MB-231细胞迁移的多糖,该功效可能是通过抑制SATB-1基因表达实现,该发现为PPS的抗乳腺癌研究提供依据。展开更多
基金Supported by the Program of National Rabbit Industrial Production Technology System of Ministry of Agriculture~~
文摘[Objective] The aim was to establish a molecular biological method for identifying coccidium species.[Method]First,the pure species of Eimeria intestinalis was isolated by using single-oocyst isolation technique.Then,according to the 18s rDNA and 5.8s rDNA sequences of Eimeria coccidia published in GenBank,a pair of specific primers was designed and synthesized to amplify the internal transcribed spacer 1（ITS-1）.Finally,the PCR products were sent for sequencing.[Result]The pure species of E.intestinalis was isolated and the result of agarose gel electrophoresis showed that the PCR product was 434 bp,and at least 27-sporulated oocysts could be detected.[Conclusion]The research will provide a basis for accurate identification of coccidium species and strains.
基金Supported by Projects for Transgenic Research (2008ZX08006-002,2008ZX08006-003,2008ZX08010-003,2008ZX08011-004)Hubei Key Laboratory Project (2011ZD127)~~
文摘[Objective] The study aimed at cloning and analyzing the insulin-like growth factor-1 （IGF-1） gene from liver of Hubei white swine. [Method] The total RNA was extracted by using Trizol from the liver of Hubei white swine and used as template to amplify IGF-1 gene cDNA by RT-PCR. The cDNA product was cloned into pCRII vector, screened with blue-white colonies, digested with double enzymes and sequenced. [Result] The sequencing result indicated that the IGF-1 gene consisted of 607 nucleotides, containing 5＇-untranslated region at nucleotides 1-145, a complete ORF at nucleotides 146-538 encoding 130 amino acids, and 3＇-untranslated region at nucleotides 539-607. It shared 100% homology with the porcine IGF-1 gene reported by Muller et al. [Conclusion] The successful cloning and sequencing of the Hubei white swine IGF-1 gene confirmed that IGF-I gene was highly conserved, which provided technical basis for the use of transgenic technology for breeding of Hubei white swine.
文摘目的分析基于活性导向分离茯苓中具有抑制人乳腺癌MDA-MB-231细胞迁移的成分并探索其分子机制。方法茯苓经醇提去除小分子成分,水提醇沉淀后得茯苓多糖(PPS),PPS经DEAE Sepharose Fast Flow阴离子交换树脂分离和Superdex-75系列凝胶纯化后活性跟踪得PPS10-2,并对PPS10-2进行高效凝胶渗透色谱(HPGPC)法分析、单糖组成测定和活性及机制探索。结果PPS抑制人乳腺癌MDA-MB-231细胞迁移,采用活性跟踪方法,从茯苓中得到活性均一多糖PPS10-2,相对分子量为5761D,主要由鼠李糖∶阿拉伯糖∶葡萄糖组成,三种单糖的摩尔比为10.0∶3.3∶7.0,PPS和PPS10-2表现出较强的抑制MDA-MB-231细胞迁移能力,并且PPS和PPS10-2均是通过抑制特异富含AT序列结合蛋白(SATB)-1的表达来抑制MDA-MB-231细胞迁移能力,推测PPS中可能是PPS10-2为主要成分发挥该作用。结论从茯苓中成功分离到具有抑制MDA-MB-231细胞迁移的多糖,该功效可能是通过抑制SATB-1基因表达实现,该发现为PPS的抗乳腺癌研究提供依据。
