Acyl-ACP thioesterases （FATs） terminates the fatty acid synthesis and allow the transport of fatty acids out of the plastids, which are the important determinants of cellular metabolism. FATB is a member of FAT enzy...Acyl-ACP thioesterases （FATs） terminates the fatty acid synthesis and allow the transport of fatty acids out of the plastids, which are the important determinants of cellular metabolism. FATB is a member of FAT enzymes that has been described previously in most of the plants. In silico cloning is a new method that utilizes the bioinformatics on the complete genome and available EST database. In this study, a full-length cDNA clone of PtFATB gene was isolated from Populus tomentosa using this approach. It is 1,450 bp in length and the open reading frame encodes a peptide of 421 amino acids. The predicted amino acid sequence shows significant homology with those from other plant species, which contain typical domains owned by FATB proteins. The transcripts of PtFATB were abundant in leaves, and less in roots detected by using semiquantitative RT-PCR. When the shoots were subjected to the stress treatments （cold, dry, NaC1） and ABA （Abscisic acid）, the expression of PtFATB was only slightly reduced under the treatment of low temperature. This suggests that the expression of PtFATB is in a constitutive fashion. This study provides the basis not only for the identification and characterization of this gene but also for the improvement of cold tolerance by controlling the expression of the PtFATB gene in trees in near future.展开更多
A novel immune-related gene was expressed in Japanese flounder （Paralichthys olivaceus） injected with Vibrio anguillarum. The complete cDNA contained a 169 bp 5＇UTR, a 336 bp open reading frame （ORF） encoding 111...A novel immune-related gene was expressed in Japanese flounder （Paralichthys olivaceus） injected with Vibrio anguillarum. The complete cDNA contained a 169 bp 5＇UTR, a 336 bp open reading frame （ORF） encoding 111 amino acids and a 556bp 3＇UTR. Six exons and five introns were identified in the PoIR2 gene. Blastp similarity comparison showed its encoding protein had 50% similarity to Danio rerio neuromedin S （NMS）, but further alignment indicated they did not have NMS C-terminal conservational signature domain. So it was not defined as an NMS homologue. Protein structure analysis indicated it had a 26aa sig- nal peptide and was a secretory pathway protein. RT-PCR demonstrated that the expression of PoIR2 was quickly induced and drastically increased in liver, kidney, spleen, gills, intestine, heart, and skeletal muscle after infected with V. anguillarurn. These results indicated that the PolR2 might play some important role in Japanese flounder immune response system. This gene was named PolR2 （P.olivaceus immune-related gene 2, GenBank accession number： EU224372）. The mature PoIR2 peptide was expressed in BL21 （DE3） pLysS using pET-32a（＋） vector and a great part of the recombinant mature peptide existed as soluble type.展开更多
The authors cloned and identified a new maize serine carboxypeptidase gene named ZmSCP from R15 inbred lines seedlings which were induced by Rhizoctonia solani AGI-IA. ZmSCP encodes a 332 amino acid protein with a pre...The authors cloned and identified a new maize serine carboxypeptidase gene named ZmSCP from R15 inbred lines seedlings which were induced by Rhizoctonia solani AGI-IA. ZmSCP encodes a 332 amino acid protein with a predicted molecular mass of 36.5 kDa and pI of 4.75. Phylogenetic analysis revealed that ZmSCP showed closer kinship with Oryza sativa and sorghum, which belong to the same evolutionary branch. Amino acid sequence analysis revealed that there are four types of amino acids in ZmSCP, the percentages of them are 43.1%, 26.9%, 13.9% and 13.1%. The authors subsequently purified the recombinant protein which expressed in Escherichia coli BL21 and analyzed its antimicrobial activities in vitro. Results showed that the recombinant protein inhibited hyphal growth of Rhizoctonia solani. The study suggests that the expression of ZmSCP is closely related to maize sheath blight resistance caused by Rhizoctonia solani. Further, the antifungal activity showed that ZmSCP may play at role in the disease resistance response.展开更多
基金This work was supported by project "Regulation of Composition and Saturation of Fatty Acid in Trees by Genetic Engineering", Introduction of Foreign Advanced Agricultural Science and Technology into China (No. 2005-4-52).
文摘Acyl-ACP thioesterases （FATs） terminates the fatty acid synthesis and allow the transport of fatty acids out of the plastids, which are the important determinants of cellular metabolism. FATB is a member of FAT enzymes that has been described previously in most of the plants. In silico cloning is a new method that utilizes the bioinformatics on the complete genome and available EST database. In this study, a full-length cDNA clone of PtFATB gene was isolated from Populus tomentosa using this approach. It is 1,450 bp in length and the open reading frame encodes a peptide of 421 amino acids. The predicted amino acid sequence shows significant homology with those from other plant species, which contain typical domains owned by FATB proteins. The transcripts of PtFATB were abundant in leaves, and less in roots detected by using semiquantitative RT-PCR. When the shoots were subjected to the stress treatments （cold, dry, NaC1） and ABA （Abscisic acid）, the expression of PtFATB was only slightly reduced under the treatment of low temperature. This suggests that the expression of PtFATB is in a constitutive fashion. This study provides the basis not only for the identification and characterization of this gene but also for the improvement of cold tolerance by controlling the expression of the PtFATB gene in trees in near future.
基金supported by grants from the National High Technology Research and Development Program of China (Nos.2006AA10A404 and 2006AA10A414)
文摘A novel immune-related gene was expressed in Japanese flounder （Paralichthys olivaceus） injected with Vibrio anguillarum. The complete cDNA contained a 169 bp 5＇UTR, a 336 bp open reading frame （ORF） encoding 111 amino acids and a 556bp 3＇UTR. Six exons and five introns were identified in the PoIR2 gene. Blastp similarity comparison showed its encoding protein had 50% similarity to Danio rerio neuromedin S （NMS）, but further alignment indicated they did not have NMS C-terminal conservational signature domain. So it was not defined as an NMS homologue. Protein structure analysis indicated it had a 26aa sig- nal peptide and was a secretory pathway protein. RT-PCR demonstrated that the expression of PoIR2 was quickly induced and drastically increased in liver, kidney, spleen, gills, intestine, heart, and skeletal muscle after infected with V. anguillarurn. These results indicated that the PolR2 might play some important role in Japanese flounder immune response system. This gene was named PolR2 （P.olivaceus immune-related gene 2, GenBank accession number： EU224372）. The mature PoIR2 peptide was expressed in BL21 （DE3） pLysS using pET-32a（＋） vector and a great part of the recombinant mature peptide existed as soluble type.
基金Acknowledgments This research was financially supported by the Natural Science Foundation （30900901）, the Science and Technology Department Application Foundation of Sichuan province （2006J13-039）, and the Agriculture Project of Ministry （2011ZX08003-003）.
文摘The authors cloned and identified a new maize serine carboxypeptidase gene named ZmSCP from R15 inbred lines seedlings which were induced by Rhizoctonia solani AGI-IA. ZmSCP encodes a 332 amino acid protein with a predicted molecular mass of 36.5 kDa and pI of 4.75. Phylogenetic analysis revealed that ZmSCP showed closer kinship with Oryza sativa and sorghum, which belong to the same evolutionary branch. Amino acid sequence analysis revealed that there are four types of amino acids in ZmSCP, the percentages of them are 43.1%, 26.9%, 13.9% and 13.1%. The authors subsequently purified the recombinant protein which expressed in Escherichia coli BL21 and analyzed its antimicrobial activities in vitro. Results showed that the recombinant protein inhibited hyphal growth of Rhizoctonia solani. The study suggests that the expression of ZmSCP is closely related to maize sheath blight resistance caused by Rhizoctonia solani. Further, the antifungal activity showed that ZmSCP may play at role in the disease resistance response.
