Objective:To determine the genetic diversity of Plasmodium(P.)knowlesi isolates from Sabah,Malaysian Borneo and Peninsular Malaysia,targeting the S-type SSU rRNA gene and including aspects of natural selection and hap...Objective:To determine the genetic diversity of Plasmodium(P.)knowlesi isolates from Sabah,Malaysian Borneo and Peninsular Malaysia,targeting the S-type SSU rRNA gene and including aspects of natural selection and haplotype.Methods:Thirty-nine blood samples infected with P.knowlesi were collected in Sabah,Malaysian Borneo and Peninsular Malaysia.The S-type SSU rRNA gene was amplified using polymerase chain reaction,cloned into a vector,and sequenced.The natural selection and haplotype of the S-type SSU rRNA gene sequences were determined using DnaSP v6 and illustrated using NETWORK v10.This study's 39 S-type SSU rRNA sequences and eight sequences from the Genbank database were subjected to phylogenetic analysis using MEGA 11.Results:Overall,the phylogenetic analysis showed no evidence of a geographical cluster of P.knowlesi isolates from different areas in Malaysia based on the S-type SSU rRNA gene sequences.The S-type SSU rRNA gene sequences were relatively conserved and with a purifying effect.Haplotype sharing of the S-type SSU rRNA gene was observed between the P.knowlesi isolates in Sabah,Malaysian Borneo,but not between Sabah,Malaysian Borneo and Peninsular Malaysia.Conclusions:This study suggests that the S-type SSU rRNA gene of P.knowlesi isolates in Sabah,Malaysian Borneo,and Peninsular Malaysia has fewer polymorphic sites,representing the conservation of the gene.These features make the S-type SSU rRNA gene suitable for comparative studies,such as determining the evolutionary relationships and common ancestry among P.knowlesi species.展开更多
Endoreticulatus bombycis is a new pathogenic microspordia isolated from the silkworm larvae. With polymerase chain reaction (PCR) method,we amplified a fragment of the core sequence of the small subunit ribosomal RNA ...Endoreticulatus bombycis is a new pathogenic microspordia isolated from the silkworm larvae. With polymerase chain reaction (PCR) method,we amplified a fragment of the core sequence of the small subunit ribosomal RNA (SSUrRNA) of Endoreticulatus bombycis. The SSUrRNA fragment was inserted into pMD18-T Vector and then cloned. It had a length of 1 230 nucleotides and a percentage GC content of 51.3%. The accession number in GenBank is gi11181769|AY009115. The secondary structure model of the SSUrRNA of Endoreticulatus bombycis was constructed both on the bases of the sequence alignment and RNAFOLD program of the PC-GENE package. The secondary structure model revealed that Endoreticulatus bombycis lacked many eukaryotic helices, such as helix10, helix 11, helix 18, helix 43 and helix 46,but it has V4 region and has more of prokaryotic character. Analyzed by Blast, Endoreticulatus bombycis, Endoreticulatus schubergi and Pleistophora sp.(Sd-Nu-IW8201) have a high similarity, over 98%. Phylogeny tree of Endoreticulatus and Pleistophora species showed that Endoreticulatus and Pleistophora sp.(Sd-Nu-IW8201) was in a group and the other Pleistophora species were in another group .展开更多
基金This study was supported by the Ministry of Higher Education,Malaysia(FRGS0322-SG-1/2013)Universiti Malaysia Sabah(GUG0521-2/2020).
文摘Objective:To determine the genetic diversity of Plasmodium(P.)knowlesi isolates from Sabah,Malaysian Borneo and Peninsular Malaysia,targeting the S-type SSU rRNA gene and including aspects of natural selection and haplotype.Methods:Thirty-nine blood samples infected with P.knowlesi were collected in Sabah,Malaysian Borneo and Peninsular Malaysia.The S-type SSU rRNA gene was amplified using polymerase chain reaction,cloned into a vector,and sequenced.The natural selection and haplotype of the S-type SSU rRNA gene sequences were determined using DnaSP v6 and illustrated using NETWORK v10.This study's 39 S-type SSU rRNA sequences and eight sequences from the Genbank database were subjected to phylogenetic analysis using MEGA 11.Results:Overall,the phylogenetic analysis showed no evidence of a geographical cluster of P.knowlesi isolates from different areas in Malaysia based on the S-type SSU rRNA gene sequences.The S-type SSU rRNA gene sequences were relatively conserved and with a purifying effect.Haplotype sharing of the S-type SSU rRNA gene was observed between the P.knowlesi isolates in Sabah,Malaysian Borneo,but not between Sabah,Malaysian Borneo and Peninsular Malaysia.Conclusions:This study suggests that the S-type SSU rRNA gene of P.knowlesi isolates in Sabah,Malaysian Borneo,and Peninsular Malaysia has fewer polymorphic sites,representing the conservation of the gene.These features make the S-type SSU rRNA gene suitable for comparative studies,such as determining the evolutionary relationships and common ancestry among P.knowlesi species.
文摘Endoreticulatus bombycis is a new pathogenic microspordia isolated from the silkworm larvae. With polymerase chain reaction (PCR) method,we amplified a fragment of the core sequence of the small subunit ribosomal RNA (SSUrRNA) of Endoreticulatus bombycis. The SSUrRNA fragment was inserted into pMD18-T Vector and then cloned. It had a length of 1 230 nucleotides and a percentage GC content of 51.3%. The accession number in GenBank is gi11181769|AY009115. The secondary structure model of the SSUrRNA of Endoreticulatus bombycis was constructed both on the bases of the sequence alignment and RNAFOLD program of the PC-GENE package. The secondary structure model revealed that Endoreticulatus bombycis lacked many eukaryotic helices, such as helix10, helix 11, helix 18, helix 43 and helix 46,but it has V4 region and has more of prokaryotic character. Analyzed by Blast, Endoreticulatus bombycis, Endoreticulatus schubergi and Pleistophora sp.(Sd-Nu-IW8201) have a high similarity, over 98%. Phylogeny tree of Endoreticulatus and Pleistophora species showed that Endoreticulatus and Pleistophora sp.(Sd-Nu-IW8201) was in a group and the other Pleistophora species were in another group .
