β-thymosins, a family of highly conserved peptides, play a vital role in wound-healing, angiogenesis,antimicrobial process and antiviral immunity. Three novel β-thymosin-repeat proteins, named mjthm4, mjthm3 and mjt...β-thymosins, a family of highly conserved peptides, play a vital role in wound-healing, angiogenesis,antimicrobial process and antiviral immunity. Three novel β-thymosin-repeat proteins, named mjthm4, mjthm3 and mjthm2, were cloned from Marsupenaeus japonicus using expressed sequence tags（EST） from suppression subtractive hybridization. The mjthm4, mjthm3 and mjthm2 c DNAs possessed open reading frames that encoded166, 128 and 90 amino acid residue polypeptides and contained four, three and two β-thymosin actin binding modules, respectively. Blast analysis demonstrated that mjthm4, mjthm3 and mjthm2 shared high homology with known invertebrate multi-repeat β-thymosins. These proteins are ubiquitously expressed in all of the examined tissues, and the transcriptional levels were highest in the intestine. Further investigation revealed that mjthm4,mjthm3 and mjthm2 were remarkably up-regulated 6 h after WSSV infection. Moreover, while mjthm4 transcriptional levels displayed no changes, mjthm3 and mjthm2 levels decreased in the virus-resistant shrimps.The results indicate that mjthm4, mjthm3 and mjthm2 are novel multi-repeat β-thymosin homologues, have a close relationship with WSSV infection, and might contribute to a better understanding of host defense and/or virus invasion interactions in shrimps.展开更多
【目的】提高免疫活性多肽人胸腺素α1(Tα1)在番茄中的表达量。【方法】根据植物偏爱密码子对Tα1基因进行优化,并将其顺式串联成四联体,同时在单体基因间插入肠激酶剪切位点基因序列,以便表达的融合蛋白可以被肠激酶切割成单体。利用...【目的】提高免疫活性多肽人胸腺素α1(Tα1)在番茄中的表达量。【方法】根据植物偏爱密码子对Tα1基因进行优化,并将其顺式串联成四联体,同时在单体基因间插入肠激酶剪切位点基因序列,以便表达的融合蛋白可以被肠激酶切割成单体。利用农杆菌介导法将单体Tα1基因和四联体Tα1基因分别转化进入番茄。【结果】两个基因转化番茄后分别得到卡那霉素抗性再生植株19株和10株,PCR和Southern杂交检测证明,部分番茄基因组中整合了外源基因。经过Western和ELISA分析,串联的Tα1基因已经在番茄果实中表达,表达量最高相当于20.2μg T α1·g-1果实鲜重,高于单体基因的表达量。【结论】基因串联的方式提高了Tα1基因在番茄中的表达量。展开更多
基金The National High Technology Research and Development Program(863 Program)of China under contract No.2012AA092205the Major State Basic Research Development Program(973 Program)of China under contract No.2012CB114403the China Agriculture Research System-47,and the Scientific Research Foundation of Third Institute of Oceanography,SOA under contract No.2011018
文摘β-thymosins, a family of highly conserved peptides, play a vital role in wound-healing, angiogenesis,antimicrobial process and antiviral immunity. Three novel β-thymosin-repeat proteins, named mjthm4, mjthm3 and mjthm2, were cloned from Marsupenaeus japonicus using expressed sequence tags（EST） from suppression subtractive hybridization. The mjthm4, mjthm3 and mjthm2 c DNAs possessed open reading frames that encoded166, 128 and 90 amino acid residue polypeptides and contained four, three and two β-thymosin actin binding modules, respectively. Blast analysis demonstrated that mjthm4, mjthm3 and mjthm2 shared high homology with known invertebrate multi-repeat β-thymosins. These proteins are ubiquitously expressed in all of the examined tissues, and the transcriptional levels were highest in the intestine. Further investigation revealed that mjthm4,mjthm3 and mjthm2 were remarkably up-regulated 6 h after WSSV infection. Moreover, while mjthm4 transcriptional levels displayed no changes, mjthm3 and mjthm2 levels decreased in the virus-resistant shrimps.The results indicate that mjthm4, mjthm3 and mjthm2 are novel multi-repeat β-thymosin homologues, have a close relationship with WSSV infection, and might contribute to a better understanding of host defense and/or virus invasion interactions in shrimps.
文摘【目的】提高免疫活性多肽人胸腺素α1(Tα1)在番茄中的表达量。【方法】根据植物偏爱密码子对Tα1基因进行优化,并将其顺式串联成四联体,同时在单体基因间插入肠激酶剪切位点基因序列,以便表达的融合蛋白可以被肠激酶切割成单体。利用农杆菌介导法将单体Tα1基因和四联体Tα1基因分别转化进入番茄。【结果】两个基因转化番茄后分别得到卡那霉素抗性再生植株19株和10株,PCR和Southern杂交检测证明,部分番茄基因组中整合了外源基因。经过Western和ELISA分析,串联的Tα1基因已经在番茄果实中表达,表达量最高相当于20.2μg T α1·g-1果实鲜重,高于单体基因的表达量。【结论】基因串联的方式提高了Tα1基因在番茄中的表达量。
