[Objective] The research aimed at cloning and analyzing mitochondrial cytochrome oxidase I gene（cox 1）of C.suppressalis.[Method] The mitochondrial cox 1 gene of C.suppressalis was cloned with PCR method and sequence...[Objective] The research aimed at cloning and analyzing mitochondrial cytochrome oxidase I gene（cox 1）of C.suppressalis.[Method] The mitochondrial cox 1 gene of C.suppressalis was cloned with PCR method and sequenced.Then,cox1 sequences of other 21 Lepidopteran species were obtained by blasting the GenBank with cox 1 gene sequence of C.suppressalis.Finally,homology comparison and molecular phylogenitic analysis among the 22 Lepidopteran species were conducted.[Result] The open reading frame of cox 1 gene from C.suppressalis contained 1 531 nucleotides encoding a putative protein of 510 amino acids.The cox1 gene used a start codon CGA,and an incomplete termination codon composed of only T.Based on the amino acid sequences of cox 1,the molecular phylogenetic tree of Lepidoptera was reconstructed using the maximum likelihood（ML）method.The molecular phylogenetic tree was similar to the morphological phylogenetic tree mainly,but also showed some differences.[Conclusion] The result will provide reference for further research on expression and application of cox 1 gene.展开更多
The whole mitochondrial genome sequence of red fox （Vuples vuples） was determined. It had a total length of 16 723 bp. As in most mammal mitochondrial genome, it contained 13 protein coding genes, two ribosome RNA g...The whole mitochondrial genome sequence of red fox （Vuples vuples） was determined. It had a total length of 16 723 bp. As in most mammal mitochondrial genome, it contained 13 protein coding genes, two ribosome RNA genes, 22 transfer RNA genes and one control region. The base composition was 31.3% A, 26.1% C, 14.8% G and 27.8% T, respectively. The codon usage of red fox, arctic fox, gray wolf, domestic dog and coyote followed the same pattern except for an unusual ATT start codon, which initiates the NADH dehydrogenase subunit 3 gene in the red fox. A long tandem repeat rich in AC was found between conserved sequence block 1 and 2 in the control region. In order to confirm the phylogenetic relationships of red fox to other canids, phylogenetic trees were reconstructed by neighbor-joining and maximum parsimony methods using 12 concatenated heavy-strand protein-coding genes. The result indicated that arctic fox was the sister group of red fox and they both belong to the red fox-like clade in family Canidae, while gray wolf, domestic dog and coyote belong to wolf-like clade. The result was in accordance with existing phylogenetic results.展开更多
Chlorophyta species are common in the southern and northern coastal areas of China. In recent years, frequent green tide incidents in Chinese coastal waters have raised concerns and attracted the attention of scientis...Chlorophyta species are common in the southern and northern coastal areas of China. In recent years, frequent green tide incidents in Chinese coastal waters have raised concerns and attracted the attention of scientists. In this paper, we sequenced the 18S rDNA genes, the internal transcribed spacer （ITS） regions and the rbcL genes in seven organisms and obtained 536-566 bp long ITS sequences, 1 377-I 407 bp long rbcL sequences and 1 718-1 761 bp long partial 18S rDNA sequences. The GC base pair content was highest in the ITS regions and lowest in the rbcL genes. The sequencing results showed that the three Ulvaprolifera （or U. pertusa） gene sequences from Qingdao and Nan＇ao Island were identical. The ITS, 18S rDNA and rbcL genes in U. prolifera and U. pertusa from different sea areas in China were unchanged by geographic distance. U.flexuosa had the least evolutionary distance from U. californica in both the ITS regions （0.009） and the 18S rDNA （0.002）. These data verified that Ulva and Enteromorpha are not separate genera.展开更多
In the present study,the partial gene sequences of P32 protein,an immunogenic envelope protein of Capripoxviruses (CaPV),were analyzed to assess the genetic relationship among sheeppox and goatpox virus isolates,and r...In the present study,the partial gene sequences of P32 protein,an immunogenic envelope protein of Capripoxviruses (CaPV),were analyzed to assess the genetic relationship among sheeppox and goatpox virus isolates,and restriction enzyme specific PCR-RFLP was developed to differentiate CaPV strains.A total of six goatpox virus (GTPV) and nine sheeppox virus (SPPV) isolates of Indian origin were included in the sequence analysis of the attachment gene.The sequence analysis revealed a high degree of sequence identity among all the Indian SPPV and GTPV isolates at both nucleotide and amino acid levels.Phylogenetic analysis showed three distinct clusters of SPPV,GTPV and Lumpy skin disease virus (LSDV) isolates.Further,multiple sequence alignment revealed a unique change at G120A in all GTPV isolates resulting in the formation of Dra I restriction site in lieu of EcoR I,which is present in SPPV isolates studied.This change was unique and exploited to develop restriction enzyme specific PCR-RFLP for detection and differentiation of SPPV and GTPV strains.The optimized PCR-RFLP was validated using a total of fourteen (n=14) cell culture isolates and twenty two (n=22) known clinical samples of CaPV.The Restriction Enzyme specific PCR-RFLP to differentiate both species will allow a rapid differential diagnosis during CaPV outbreaks particularly in mixed flocks of sheep and goats and could be an adjunct/supportive tool for complete gene or virus genome sequencing methods.展开更多
基金Supported by New Century Program for Excellent Talents of Ministry of Education of China(NCET-07-0251)Talents Foundation of Anhui Province(08040106803)~~
文摘[Objective] The research aimed at cloning and analyzing mitochondrial cytochrome oxidase I gene（cox 1）of C.suppressalis.[Method] The mitochondrial cox 1 gene of C.suppressalis was cloned with PCR method and sequenced.Then,cox1 sequences of other 21 Lepidopteran species were obtained by blasting the GenBank with cox 1 gene sequence of C.suppressalis.Finally,homology comparison and molecular phylogenitic analysis among the 22 Lepidopteran species were conducted.[Result] The open reading frame of cox 1 gene from C.suppressalis contained 1 531 nucleotides encoding a putative protein of 510 amino acids.The cox1 gene used a start codon CGA,and an incomplete termination codon composed of only T.Based on the amino acid sequences of cox 1,the molecular phylogenetic tree of Lepidoptera was reconstructed using the maximum likelihood（ML）method.The molecular phylogenetic tree was similar to the morphological phylogenetic tree mainly,but also showed some differences.[Conclusion] The result will provide reference for further research on expression and application of cox 1 gene.
基金National Natural Science Foundation of China (30370218)the program for New Century Excellent Talents in University (NCET-07-0507)+1 种基金 the Project of Science and Technology Development Plan in Shandong Province (2007GG2009011)Shandong Science Fund for Distinguished Young Scholars (2005BS02005)
文摘The whole mitochondrial genome sequence of red fox （Vuples vuples） was determined. It had a total length of 16 723 bp. As in most mammal mitochondrial genome, it contained 13 protein coding genes, two ribosome RNA genes, 22 transfer RNA genes and one control region. The base composition was 31.3% A, 26.1% C, 14.8% G and 27.8% T, respectively. The codon usage of red fox, arctic fox, gray wolf, domestic dog and coyote followed the same pattern except for an unusual ATT start codon, which initiates the NADH dehydrogenase subunit 3 gene in the red fox. A long tandem repeat rich in AC was found between conserved sequence block 1 and 2 in the control region. In order to confirm the phylogenetic relationships of red fox to other canids, phylogenetic trees were reconstructed by neighbor-joining and maximum parsimony methods using 12 concatenated heavy-strand protein-coding genes. The result indicated that arctic fox was the sister group of red fox and they both belong to the red fox-like clade in family Canidae, while gray wolf, domestic dog and coyote belong to wolf-like clade. The result was in accordance with existing phylogenetic results.
基金Supported by the National Natural Science Foundation of China (No.30570125)the Key Construction Laboratory of Marine Biotechnology of Jiangsu Province (No. 2010HS03)
文摘Chlorophyta species are common in the southern and northern coastal areas of China. In recent years, frequent green tide incidents in Chinese coastal waters have raised concerns and attracted the attention of scientists. In this paper, we sequenced the 18S rDNA genes, the internal transcribed spacer （ITS） regions and the rbcL genes in seven organisms and obtained 536-566 bp long ITS sequences, 1 377-I 407 bp long rbcL sequences and 1 718-1 761 bp long partial 18S rDNA sequences. The GC base pair content was highest in the ITS regions and lowest in the rbcL genes. The sequencing results showed that the three Ulvaprolifera （or U. pertusa） gene sequences from Qingdao and Nan＇ao Island were identical. The ITS, 18S rDNA and rbcL genes in U. prolifera and U. pertusa from different sea areas in China were unchanged by geographic distance. U.flexuosa had the least evolutionary distance from U. californica in both the ITS regions （0.009） and the 18S rDNA （0.002）. These data verified that Ulva and Enteromorpha are not separate genera.
文摘In the present study,the partial gene sequences of P32 protein,an immunogenic envelope protein of Capripoxviruses (CaPV),were analyzed to assess the genetic relationship among sheeppox and goatpox virus isolates,and restriction enzyme specific PCR-RFLP was developed to differentiate CaPV strains.A total of six goatpox virus (GTPV) and nine sheeppox virus (SPPV) isolates of Indian origin were included in the sequence analysis of the attachment gene.The sequence analysis revealed a high degree of sequence identity among all the Indian SPPV and GTPV isolates at both nucleotide and amino acid levels.Phylogenetic analysis showed three distinct clusters of SPPV,GTPV and Lumpy skin disease virus (LSDV) isolates.Further,multiple sequence alignment revealed a unique change at G120A in all GTPV isolates resulting in the formation of Dra I restriction site in lieu of EcoR I,which is present in SPPV isolates studied.This change was unique and exploited to develop restriction enzyme specific PCR-RFLP for detection and differentiation of SPPV and GTPV strains.The optimized PCR-RFLP was validated using a total of fourteen (n=14) cell culture isolates and twenty two (n=22) known clinical samples of CaPV.The Restriction Enzyme specific PCR-RFLP to differentiate both species will allow a rapid differential diagnosis during CaPV outbreaks particularly in mixed flocks of sheep and goats and could be an adjunct/supportive tool for complete gene or virus genome sequencing methods.
