Ubiquitin, a highly conserved stress-related protein, is assigned multiple functions, such as DNA processing, protein degradation, and ribosome synthesis. The Crassostrea hongkongensis ubiquitin gene (designated ChUbL...Ubiquitin, a highly conserved stress-related protein, is assigned multiple functions, such as DNA processing, protein degradation, and ribosome synthesis. The Crassostrea hongkongensis ubiquitin gene (designated ChUbL40) was cloned by a combination of suppressive subtractive hybridization (SSH) and rapid amplification of cDNA ends (RACE). The full-length cDNA of ChUbL40 is 496 bp in length, consisting of a 5' untranslated region (UTR) of 34 bp, a 3'-UTR of 75 bp and an open reading frame of 387 bp encoding a ubiquitin fusion protein of 128 amino acids. Analysis of the amino acid sequence of ChUbL40 reveals that UbL40 is highly conservative during evolution. The expression patterns of ChUbL40 gene in various tissues were examined by real-time PCR. The expression level of ChUbL40 in haemocytes is down-regulated at 4 h and gradually returned to its original level from 6 h to 24 h after Vibrio alginolyticus challenge. Our results suggest that ChUbL40 is ubiquitously expressed and plays an important role in immune defense against bacterial challenge.展开更多
Ubiquitin (UBI) plays a very important role in regulated non-lysosoma l ATP dependent protein degradation. In the present work, the coding sequence of Spodoptera litura UBI gene was isolated (GenBank Accession No. AF4...Ubiquitin (UBI) plays a very important role in regulated non-lysosoma l ATP dependent protein degradation. In the present work, the coding sequence of Spodoptera litura UBI gene was isolated (GenBank Accession No. AF436066). Th e le ngth of this ORF is 228bp, encoding a protein with Mr of 8.56 kD and isoelectri c point of 6.56. Multiple sequence alignment indicated that S.litura UBI is very similar to the homologous proteins of other eukaryotic species and it has 84% id entity with S.litura nucleopolyhedrovirus (SpltMNPV) UBI at amino acid level . RT -PCR analysis showed that S.litura UBI gene is ubiquitously expressed in la rva t issues which are susceptible to SpltMNPV infection. By constructing E.coli e xpre ssion vector, S.litura UBI was highly expressed and the recombinant protein was purified using Ni-NTA resin column, and currently further study on the function of S.litura UBI in SpltMNPV infection is underway.展开更多
基金Supported by the National Basic Research Program of China (973 Program) (No.2010CB126404)the CAS/SAFEA International Partnership Program for Creative Research Teams (No.KZCX2-YW-T001)the Research Program for Young Scientists at SCSIO
文摘Ubiquitin, a highly conserved stress-related protein, is assigned multiple functions, such as DNA processing, protein degradation, and ribosome synthesis. The Crassostrea hongkongensis ubiquitin gene (designated ChUbL40) was cloned by a combination of suppressive subtractive hybridization (SSH) and rapid amplification of cDNA ends (RACE). The full-length cDNA of ChUbL40 is 496 bp in length, consisting of a 5' untranslated region (UTR) of 34 bp, a 3'-UTR of 75 bp and an open reading frame of 387 bp encoding a ubiquitin fusion protein of 128 amino acids. Analysis of the amino acid sequence of ChUbL40 reveals that UbL40 is highly conservative during evolution. The expression patterns of ChUbL40 gene in various tissues were examined by real-time PCR. The expression level of ChUbL40 in haemocytes is down-regulated at 4 h and gradually returned to its original level from 6 h to 24 h after Vibrio alginolyticus challenge. Our results suggest that ChUbL40 is ubiquitously expressed and plays an important role in immune defense against bacterial challenge.
文摘Ubiquitin (UBI) plays a very important role in regulated non-lysosoma l ATP dependent protein degradation. In the present work, the coding sequence of Spodoptera litura UBI gene was isolated (GenBank Accession No. AF436066). Th e le ngth of this ORF is 228bp, encoding a protein with Mr of 8.56 kD and isoelectri c point of 6.56. Multiple sequence alignment indicated that S.litura UBI is very similar to the homologous proteins of other eukaryotic species and it has 84% id entity with S.litura nucleopolyhedrovirus (SpltMNPV) UBI at amino acid level . RT -PCR analysis showed that S.litura UBI gene is ubiquitously expressed in la rva t issues which are susceptible to SpltMNPV infection. By constructing E.coli e xpre ssion vector, S.litura UBI was highly expressed and the recombinant protein was purified using Ni-NTA resin column, and currently further study on the function of S.litura UBI in SpltMNPV infection is underway.
