Nine strains resistant to five fluoroquinolones (Ciprofloxacin, Ofloxacin, Enrofloxacin, Danofloxacin, Sarafloxacin) were isolated from clinical samples and extracted the chromosomal DNA of these strains. Designed pri...Nine strains resistant to five fluoroquinolones (Ciprofloxacin, Ofloxacin, Enrofloxacin, Danofloxacin, Sarafloxacin) were isolated from clinical samples and extracted the chromosomal DNA of these strains. Designed primers to amplify the Quinolone-resistance-determining region(QRDR) of gyrB gene, then the PCR products were cloned and the sequence was analyzed. In comparison with the standarded strain NCTC5776, no mutation was found in the QRDR of gyrB gene of all resistant strains. The result indicated that the QRDR of gyrB has little relationship with fluoroquinolone resistance to salmonella.展开更多
In this paper,a 1,860 bp sequence in IRs region of duck enteritis virus(DEV) was amplified by single oligonucleotide nested PCR with a single primer designed according to partial sequence of US1 and then a pair of pri...In this paper,a 1,860 bp sequence in IRs region of duck enteritis virus(DEV) was amplified by single oligonucleotide nested PCR with a single primer designed according to partial sequence of US1 and then a pair of primers designed according to the 3' UTR of US8 gene and 5' end of the new getting sequence were used to amplify a 2,426 bp sequence toward the TRs region.Sequence analysis revealed that the both sequences contained an identical 990 bp open reading frame of DEV US1 gene.The two ORFs were in opposite transcription orientation.Sequence comparison of the nucleotide sequence and the deduced amino acid sequence of US1 gene showed relatively high identity to Mardivirus.Phylogenetic tree analysis showed that the eleven herpesviruses viruses were classified into three groups,and the duck enteritis virus was most closely related to Mardivirus.展开更多
Degenerate primers were designed based on the conserved sequences of the Actin gene from other plants. Total RNA was extracted from the leaves of lris lacteal var.chinensis Fisch.Koidz. Actin gene fragment was obtaine...Degenerate primers were designed based on the conserved sequences of the Actin gene from other plants. Total RNA was extracted from the leaves of lris lacteal var.chinensis Fisch.Koidz. Actin gene fragment was obtained by reverse transcription polymerase chain reaction （RT-PCR） and cloned into pMD18-T vector. The positive clone identified by PCR was sequenced. The sequencing result showed that the Actin gene fragment from lris lacteal var.chinensis Fisch.Koidz contained about 598 bp, encoding 199 amino acids. Homology comparison with Actin gene sequences of other plants in the GenBank showed that it shared over 82% nueleotide sequence homology and 90% amino acid sequence homology. It indicated that this was the Actin gene. Because of the stability expression ofActin gene, it usually cited as the internal reference to study the expression and regulation of foundation in other genes of lris lacteal var.chinensis Fisch.Koidz well.展开更多
By means of PCR,the gene encoding gD of bovine herpesvirus-1 (BHV-1) strain Luojing was amplified,cloned and sequenced.The nucleotide sequence of this gD gene was (1 251 bp,)encoding 417 amino acids.Comparied with the...By means of PCR,the gene encoding gD of bovine herpesvirus-1 (BHV-1) strain Luojing was amplified,cloned and sequenced.The nucleotide sequence of this gD gene was (1 251 bp,)encoding 417 amino acids.Comparied with the published P8-2 strain,the homology of the necleotide sequence is 99.92%,and that of the deduced amino acid sequence is 100%.The results indicated that gD of BHV-1 was highly conservative.展开更多
At the early stage,the transcriptome sequencing technique was used to detect the differentially expressed gene CsFK111 between vine cucumber and dwarf cucumber D0462.The gene was cloned,and bioinformatics software too...At the early stage,the transcriptome sequencing technique was used to detect the differentially expressed gene CsFK111 between vine cucumber and dwarf cucumber D0462.The gene was cloned,and bioinformatics software tools were used to analyze and predict the gene family and this gene.There were 30 members of the cucumber F-box gene family.The coding region of the cucumber CsFK111 gene was full-length 1314 bp,which encoded 437 amino acids and was predicted to be located in the nucleus.The protein encoded by this gene was a non-transmembrane protein,and the prediction of the secondary structure showed thatβ-lamellar structure and irregular crimp were dominant.A comparison of the phylogenetic tree showed that it was closest to cantaloupe and belonged to the same branch.The results provided a basis for future study on the regulation mechanism of the CsFK111 gene on cucumber dwarfing and also laid a foundation for further study of FBK family proteins.展开更多
Chitin is the most widespread amino polysaccharide in nature. Chitin synthase （CHS） plays an important role in chitin formation in the cuticle and the peritrophic membrane （PM） lining the midgut. Total RNA was iso...Chitin is the most widespread amino polysaccharide in nature. Chitin synthase （CHS） plays an important role in chitin formation in the cuticle and the peritrophic membrane （PM） lining the midgut. Total RNA was isolated from the cuticle of Mamestra brassicae （L.） fourth instar larva, cDNA sequence was cloned by RT-PCR and Rapid Amplification of cDNA Ends （RACE）. cDNA 5 220 bp in length, contained an open reading frame of 4 704 bp coding for a polypeptide of 1 567 amino acid residues with a predicted molecular weight of 178.3 ku and its pI was 6.42. The deduced amino acid sequence from Mi brassicae （L.） shared the high level of identity with chitin synthase sequences from other insects, especially lepidopteran insects, cDNA sequence has been deposited with GenBank under accession No. GQ281761展开更多
The full-length Mlo gene was obtained by reverse transcription polymerase chain reaction （RT-PCR） and RACE. The result of sequence analysis indicated that M/o gene from Pericallis hybrida B. Nord. contained about 12...The full-length Mlo gene was obtained by reverse transcription polymerase chain reaction （RT-PCR） and RACE. The result of sequence analysis indicated that M/o gene from Pericallis hybrida B. Nord. contained about 1296bp open reading frame and encoded 431 amino acids. According to the comparison of the exogenous gene sequences by BLAST analysis and phylogenetic analysis, Mlo gene shared over 85% nucleotide homology and 98% amino acid homology. Finally, through semi-quantitative-PCR and fluorescence quantitative analysis, we found that Mlo gene showed the highest expression levels in leaves and the lowest in roots after inoculated with powdery mildew pathogen for different days.展开更多
S-adenosylmethionine （SAM） plays important role in trans-methyl reactions. Under the condition of drought （30% PEG）, salinity （200 mmol· L^-1 NaCl） and low temperature （4℃）, total RNA was extracted from ...S-adenosylmethionine （SAM） plays important role in trans-methyl reactions. Under the condition of drought （30% PEG）, salinity （200 mmol· L^-1 NaCl） and low temperature （4℃）, total RNA was extracted from the leaf and the first strand of cDNA was synthesized with reverse transcription. S-adenosylmethionine synthetase gene （SAMS gene） was amplified by PCR with the first strand cDNA as template and a pair of primers which was based on constructed ESTs sequence. Full-length SAMS gene sequence was obtained by BLAST comparison. According to the analysis, completed sequence of SAMS gene was integrality. The sequence of the SAMS gene was 1 185 bp in length with an opening reading frame （ORF） encoding 394 amino acids. The cDNA sequence showed a significant homology to the SAM genes from Phaseolus lunatus （89%）, Medicago sativa （85%）. A prokaryotic expression vectors based on pET-32b had been constructed and prokaryotic expression was analyzed in order to lay a strong foundation for resist adversity function analysis through situation of genic expression analysis.展开更多
The zinc finger proteins belong to the largest family of transcription factors.But there is little research of Cys2/His2 type zinc finger proteins in cotton,and there is no submission of correlating
The sucrose non-fermenting-1 related protein kinase（SnRK）, whose expression is induced by kinds of hyperosmotic stresses, plays a key role in improving stress resistance of plants. In order to investigate the molecu...The sucrose non-fermenting-1 related protein kinase（SnRK）, whose expression is induced by kinds of hyperosmotic stresses, plays a key role in improving stress resistance of plants. In order to investigate the molecular mechanism of low nitrogen resistance in cucumber, the full-length cDNA of SnRK gene was cloned in this study. The result showed that SnRK gene was 1 548 bp in length, encoded 515 amino acids, and had more than 80% homology with other crops. The protein encoded by this gene was an unstable and hydrophilic protein with no transmembrane structure and no signal peptide. Under nitrogen-free conditions and low nitrogen conditions, the expression pattern analysis of SnRK gene showed that this gene was up-regulated and its expression increased and was significantly higher than the normal level as the nitrogen concentration decreased. In addition, the expression of SnRK gene was also inhibited in the high nitrogen level and was significantly lower than the normal level. The result of this study would help us understand the molecular mechanism of low nitrogen resistance in cucumber.展开更多
辣椒作为常异花授粉作物,在生产实际中利用核不育两用系进行制种是理想手段。α-微管蛋白(α-tubulin)是细胞骨架的主要成分,与植物育性相关。以辣椒核不育两用系AB114为材料,采用扩增片段长度多态性(amplified fragment length polymor...辣椒作为常异花授粉作物,在生产实际中利用核不育两用系进行制种是理想手段。α-微管蛋白(α-tubulin)是细胞骨架的主要成分,与植物育性相关。以辣椒核不育两用系AB114为材料,采用扩增片段长度多态性(amplified fragment length polymorphism,AFLP)技术得到蕾期基因表达差异片段,利用cDNA末端快速扩增(rapid-amplifi cation of cDNA ends,RACE)技术克隆AFLP-F-57全长。ORF finder分析显示,该基因具有1350 bp的最大开放阅读框,编码450个氨基酸;经Standard Protein BLAST和NCBI Conserved Domain Search鉴定,该基因编码蛋白与烟草、辣椒的α-微管蛋白具有高度相似性,均含有α-tubulin保守结构域,故将其命名为CaTUA基因;RT-PCR分析表明,与可育株相比,不育株中该基因的表达量随花蕾发育呈现下降趋势,于大花蕾时期差异显著;系统进化树分析发现CaTUA蛋白与烟草TUA蛋白序列的进化距离较近。研究初步分析了CaTUA基因的功能,为进一步揭示辣椒核不育与CaTUA的关系提供了理论依据。展开更多
文摘Nine strains resistant to five fluoroquinolones (Ciprofloxacin, Ofloxacin, Enrofloxacin, Danofloxacin, Sarafloxacin) were isolated from clinical samples and extracted the chromosomal DNA of these strains. Designed primers to amplify the Quinolone-resistance-determining region(QRDR) of gyrB gene, then the PCR products were cloned and the sequence was analyzed. In comparison with the standarded strain NCTC5776, no mutation was found in the QRDR of gyrB gene of all resistant strains. The result indicated that the QRDR of gyrB has little relationship with fluoroquinolone resistance to salmonella.
基金Founding of Heilongjiang province education department major program(10541Z004)Heilongjiang province key scientific and technological program(GB04B504)
文摘In this paper,a 1,860 bp sequence in IRs region of duck enteritis virus(DEV) was amplified by single oligonucleotide nested PCR with a single primer designed according to partial sequence of US1 and then a pair of primers designed according to the 3' UTR of US8 gene and 5' end of the new getting sequence were used to amplify a 2,426 bp sequence toward the TRs region.Sequence analysis revealed that the both sequences contained an identical 990 bp open reading frame of DEV US1 gene.The two ORFs were in opposite transcription orientation.Sequence comparison of the nucleotide sequence and the deduced amino acid sequence of US1 gene showed relatively high identity to Mardivirus.Phylogenetic tree analysis showed that the eleven herpesviruses viruses were classified into three groups,and the duck enteritis virus was most closely related to Mardivirus.
基金Supported by the Postdoctoral Scientific Research Foundation of Heilongjiang Province(LBH-Q10144)the Natural Science Foundation of Heilongjiang Province(C201112)Northeast Agricultural University Doctoral Research Fund(200830)
文摘Degenerate primers were designed based on the conserved sequences of the Actin gene from other plants. Total RNA was extracted from the leaves of lris lacteal var.chinensis Fisch.Koidz. Actin gene fragment was obtained by reverse transcription polymerase chain reaction （RT-PCR） and cloned into pMD18-T vector. The positive clone identified by PCR was sequenced. The sequencing result showed that the Actin gene fragment from lris lacteal var.chinensis Fisch.Koidz contained about 598 bp, encoding 199 amino acids. Homology comparison with Actin gene sequences of other plants in the GenBank showed that it shared over 82% nueleotide sequence homology and 90% amino acid sequence homology. It indicated that this was the Actin gene. Because of the stability expression ofActin gene, it usually cited as the internal reference to study the expression and regulation of foundation in other genes of lris lacteal var.chinensis Fisch.Koidz well.
文摘By means of PCR,the gene encoding gD of bovine herpesvirus-1 (BHV-1) strain Luojing was amplified,cloned and sequenced.The nucleotide sequence of this gD gene was (1 251 bp,)encoding 417 amino acids.Comparied with the published P8-2 strain,the homology of the necleotide sequence is 99.92%,and that of the deduced amino acid sequence is 100%.The results indicated that gD of BHV-1 was highly conservative.
基金Supported by the National Natural Science Foundation of China(32272724)the National Science Foundation of Heilongjiang Province,China(LH2019C033)。
文摘At the early stage,the transcriptome sequencing technique was used to detect the differentially expressed gene CsFK111 between vine cucumber and dwarf cucumber D0462.The gene was cloned,and bioinformatics software tools were used to analyze and predict the gene family and this gene.There were 30 members of the cucumber F-box gene family.The coding region of the cucumber CsFK111 gene was full-length 1314 bp,which encoded 437 amino acids and was predicted to be located in the nucleus.The protein encoded by this gene was a non-transmembrane protein,and the prediction of the secondary structure showed thatβ-lamellar structure and irregular crimp were dominant.A comparison of the phylogenetic tree showed that it was closest to cantaloupe and belonged to the same branch.The results provided a basis for future study on the regulation mechanism of the CsFK111 gene on cucumber dwarfing and also laid a foundation for further study of FBK family proteins.
基金Supported by Natural Science Foundation of Heilongjiang Province (C2007-7)Scientific and Technical Innovation Fund of Harbin (RC2006QN002027)Northeast Agricultural University Research Fund (2005)
文摘Chitin is the most widespread amino polysaccharide in nature. Chitin synthase （CHS） plays an important role in chitin formation in the cuticle and the peritrophic membrane （PM） lining the midgut. Total RNA was isolated from the cuticle of Mamestra brassicae （L.） fourth instar larva, cDNA sequence was cloned by RT-PCR and Rapid Amplification of cDNA Ends （RACE）. cDNA 5 220 bp in length, contained an open reading frame of 4 704 bp coding for a polypeptide of 1 567 amino acid residues with a predicted molecular weight of 178.3 ku and its pI was 6.42. The deduced amino acid sequence from Mi brassicae （L.） shared the high level of identity with chitin synthase sequences from other insects, especially lepidopteran insects, cDNA sequence has been deposited with GenBank under accession No. GQ281761
基金Supported by Postdoctoral Scientifi c Research Foundation of Heilongjiang Province(LBH-Q10144)the Natural Science Foundation of Heilongjiang Province(C201112)Northeast Agricultural University Doctoral Research Fund(200830)
文摘The full-length Mlo gene was obtained by reverse transcription polymerase chain reaction （RT-PCR） and RACE. The result of sequence analysis indicated that M/o gene from Pericallis hybrida B. Nord. contained about 1296bp open reading frame and encoded 431 amino acids. According to the comparison of the exogenous gene sequences by BLAST analysis and phylogenetic analysis, Mlo gene shared over 85% nucleotide homology and 98% amino acid homology. Finally, through semi-quantitative-PCR and fluorescence quantitative analysis, we found that Mlo gene showed the highest expression levels in leaves and the lowest in roots after inoculated with powdery mildew pathogen for different days.
基金National Science Foundation (30570990)Heilongjiang Province Educational Committee Science Research Foundation (11521023)
文摘S-adenosylmethionine （SAM） plays important role in trans-methyl reactions. Under the condition of drought （30% PEG）, salinity （200 mmol· L^-1 NaCl） and low temperature （4℃）, total RNA was extracted from the leaf and the first strand of cDNA was synthesized with reverse transcription. S-adenosylmethionine synthetase gene （SAMS gene） was amplified by PCR with the first strand cDNA as template and a pair of primers which was based on constructed ESTs sequence. Full-length SAMS gene sequence was obtained by BLAST comparison. According to the analysis, completed sequence of SAMS gene was integrality. The sequence of the SAMS gene was 1 185 bp in length with an opening reading frame （ORF） encoding 394 amino acids. The cDNA sequence showed a significant homology to the SAM genes from Phaseolus lunatus （89%）, Medicago sativa （85%）. A prokaryotic expression vectors based on pET-32b had been constructed and prokaryotic expression was analyzed in order to lay a strong foundation for resist adversity function analysis through situation of genic expression analysis.
文摘The zinc finger proteins belong to the largest family of transcription factors.But there is little research of Cys2/His2 type zinc finger proteins in cotton,and there is no submission of correlating
基金Supported by the National Natural Science Foundation of China(31101545)the Planning Subject of Twelfth Five-year-plan in National Science and Technology for Rural Development in China(2012AA100105)
文摘The sucrose non-fermenting-1 related protein kinase（SnRK）, whose expression is induced by kinds of hyperosmotic stresses, plays a key role in improving stress resistance of plants. In order to investigate the molecular mechanism of low nitrogen resistance in cucumber, the full-length cDNA of SnRK gene was cloned in this study. The result showed that SnRK gene was 1 548 bp in length, encoded 515 amino acids, and had more than 80% homology with other crops. The protein encoded by this gene was an unstable and hydrophilic protein with no transmembrane structure and no signal peptide. Under nitrogen-free conditions and low nitrogen conditions, the expression pattern analysis of SnRK gene showed that this gene was up-regulated and its expression increased and was significantly higher than the normal level as the nitrogen concentration decreased. In addition, the expression of SnRK gene was also inhibited in the high nitrogen level and was significantly lower than the normal level. The result of this study would help us understand the molecular mechanism of low nitrogen resistance in cucumber.
文摘辣椒作为常异花授粉作物,在生产实际中利用核不育两用系进行制种是理想手段。α-微管蛋白(α-tubulin)是细胞骨架的主要成分,与植物育性相关。以辣椒核不育两用系AB114为材料,采用扩增片段长度多态性(amplified fragment length polymorphism,AFLP)技术得到蕾期基因表达差异片段,利用cDNA末端快速扩增(rapid-amplifi cation of cDNA ends,RACE)技术克隆AFLP-F-57全长。ORF finder分析显示,该基因具有1350 bp的最大开放阅读框,编码450个氨基酸;经Standard Protein BLAST和NCBI Conserved Domain Search鉴定,该基因编码蛋白与烟草、辣椒的α-微管蛋白具有高度相似性,均含有α-tubulin保守结构域,故将其命名为CaTUA基因;RT-PCR分析表明,与可育株相比,不育株中该基因的表达量随花蕾发育呈现下降趋势,于大花蕾时期差异显著;系统进化树分析发现CaTUA蛋白与烟草TUA蛋白序列的进化距离较近。研究初步分析了CaTUA基因的功能,为进一步揭示辣椒核不育与CaTUA的关系提供了理论依据。
