[Objective] The aim was to lay a foundation for research of contraceptive rodenticide with LDH-C4 as target protein. [ Method] EST sequence of LDH-C gene from black-lipped pika was cloned by PCR with degenerate primer...[Objective] The aim was to lay a foundation for research of contraceptive rodenticide with LDH-C4 as target protein. [ Method] EST sequence of LDH-C gene from black-lipped pika was cloned by PCR with degenerate primers; then the full length open reading frame (ORF) and 3'UTR sequence were cloned by RACE technique. [ Result] The full length cDNA was 1 498 bp containing an ORF of 996 bp and a 3'UTR of 486 bp. The ORF encoded a polypeptide of 332 amino acids. The alignment of LDH-C gene ORF nucleotide sequences from different species showed that the gene was conserved even between large taxons. The phylogenic tree showed that black-lipped pika LDH-C was closer to prima- tes and artiodactyla than to rodents. [Conclusion] cDNA sequence of LDH-C gene from black-lipped pika was cloned successfully.展开更多
Aldehyde dehydrogenases(ALDHs) represent a large protein family, which includes several members that catalyze the oxidation of an aldehyde to its corresponding carboxylic acid in plants. Genes encoding members of th...Aldehyde dehydrogenases(ALDHs) represent a large protein family, which includes several members that catalyze the oxidation of an aldehyde to its corresponding carboxylic acid in plants. Genes encoding members of the ALDH7 subfamily have been suggested to play important roles in various stress adaptations in plants. In this study, quantitative RT-PCR analysis revealed that a maize ALDH7 subfamily member(ZmALDH7B6) was constitutively expressed in various organs, including roots, leaves, immature ears, tassels, and developing seeds. The abundance of ZmALDH7B6 mRNA transcripts in maize roots was increased by ammonium, NaCl, and mannitol treatments. To further analyze tissue-specific and stress-induced expression patterns, the 1.5-kb 5′-flanking ZmALDH7B6 promoter region was fused to the β-glucuronidase(GUS) reporter gene and introduced into maize plants. In roots of independent transgenic lines, there was significant induction of GUS activity in response to ammonium supply, confirming ammonium-dependent expression of ZmALDH7B6 at the transcript level. Histochemical staining showed that GUS activity driven by the ZmALDH7B6 promoter was mainly localized in the vascular tissues of maize roots. These results suggested that ZmALDH7B6 is induced by multiple environmental stresses in maize roots, and may play a role in detoxifying aldehydes, particularly in vascular tissue.展开更多
Low-ion-density discontinuous polyacrylamide electrophoresis (pH 8.0) of LDH from different tissuse of four species (Tetrastes bonasia,Chrysolophus pictus, Phasians colchicus, Gallus gallus domesticus) in Galliformes...Low-ion-density discontinuous polyacrylamide electrophoresis (pH 8.0) of LDH from different tissuse of four species (Tetrastes bonasia,Chrysolophus pictus, Phasians colchicus, Gallus gallus domesticus) in Galliformes showed that this electrophoresis system was suitable for analysing avian LDH. Thermostabilty to heat and nonsusceptibility to urea inhibition of low-density urea of the four liver LDHs were in such order as LDH1> LDH2> LDH3> LDH4> LDH5. And it appeared. by comparing relative movable ratio of A and B subunits of LDHs, that difference exists between Phasianidae and Tetraonidae and in Phasianidae Phasianus was closer to Chrysolophus than to Gallus.展开更多
基金Supported by State Ethnic Affairs Commission of P.R.C.(08XN04)Applicable and Fundamental Research Funds of SichuanProvince(2008JY0068)Academic Culture and TechnologyLeaders in Sichuan Province Foundation~~
文摘[Objective] The aim was to lay a foundation for research of contraceptive rodenticide with LDH-C4 as target protein. [ Method] EST sequence of LDH-C gene from black-lipped pika was cloned by PCR with degenerate primers; then the full length open reading frame (ORF) and 3'UTR sequence were cloned by RACE technique. [ Result] The full length cDNA was 1 498 bp containing an ORF of 996 bp and a 3'UTR of 486 bp. The ORF encoded a polypeptide of 332 amino acids. The alignment of LDH-C gene ORF nucleotide sequences from different species showed that the gene was conserved even between large taxons. The phylogenic tree showed that black-lipped pika LDH-C was closer to prima- tes and artiodactyla than to rodents. [Conclusion] cDNA sequence of LDH-C gene from black-lipped pika was cloned successfully.
基金financially supported by the National 863 Program of China(2012AA100306)the National 973 Program of China(2011CB100305)+1 种基金the National Natural Science Foundation of China(30971863,31121062)the Ministry of Agriculture of China(2011ZX08003-005)
文摘Aldehyde dehydrogenases(ALDHs) represent a large protein family, which includes several members that catalyze the oxidation of an aldehyde to its corresponding carboxylic acid in plants. Genes encoding members of the ALDH7 subfamily have been suggested to play important roles in various stress adaptations in plants. In this study, quantitative RT-PCR analysis revealed that a maize ALDH7 subfamily member(ZmALDH7B6) was constitutively expressed in various organs, including roots, leaves, immature ears, tassels, and developing seeds. The abundance of ZmALDH7B6 mRNA transcripts in maize roots was increased by ammonium, NaCl, and mannitol treatments. To further analyze tissue-specific and stress-induced expression patterns, the 1.5-kb 5′-flanking ZmALDH7B6 promoter region was fused to the β-glucuronidase(GUS) reporter gene and introduced into maize plants. In roots of independent transgenic lines, there was significant induction of GUS activity in response to ammonium supply, confirming ammonium-dependent expression of ZmALDH7B6 at the transcript level. Histochemical staining showed that GUS activity driven by the ZmALDH7B6 promoter was mainly localized in the vascular tissues of maize roots. These results suggested that ZmALDH7B6 is induced by multiple environmental stresses in maize roots, and may play a role in detoxifying aldehydes, particularly in vascular tissue.
文摘Low-ion-density discontinuous polyacrylamide electrophoresis (pH 8.0) of LDH from different tissuse of four species (Tetrastes bonasia,Chrysolophus pictus, Phasians colchicus, Gallus gallus domesticus) in Galliformes showed that this electrophoresis system was suitable for analysing avian LDH. Thermostabilty to heat and nonsusceptibility to urea inhibition of low-density urea of the four liver LDHs were in such order as LDH1> LDH2> LDH3> LDH4> LDH5. And it appeared. by comparing relative movable ratio of A and B subunits of LDHs, that difference exists between Phasianidae and Tetraonidae and in Phasianidae Phasianus was closer to Chrysolophus than to Gallus.