ABSTRACT
<p><b>OBJECTIVE</b>To explore molecular and genetic mechanism of 3 cases of para-Bombay blood group.</p><p><b>METHODS</b>The bood samples of proband and family members were selected to identify their blood groups with conventional serologic methods, and salivary components carrying the ABH antigens were detected. The coding regions of FUT1 as well as exon 6 and 7 of the ABO gene were amplified using polymerase chain reaction(PCR), and the FUT1 gene was directly sequenced.</p><p><b>RESULTS</b>All the 3 cases of proband were confirmed as para-Bombay blood group. Direct sequencing revealed h new2 (nt328G→A) and h1(nt 547 ΔAG) in FUT1 gene of the proband 1, and FUT1 genotype was h1/h new2. However, the genotypes of his parents were H/h1 and H/h new2, which were non-Bombay individuals. The FUT1 genotypes of proband 2 and 3 were h1h2 (nt 547 ΔAG) and h1h2 (nt 880 ΔTT), respectively.</p><p><b>CONCLUSION</b>The technology of molecular biology can be used to detect the base deletion mutations in FUT1 gene, which contributes to the analysis of molecular and genetic mechanism of para-Bombay blood group.</p>
ABSTRACT
In order to study the molecular characterization of the hantavirus isolated from Apodemus peninsulae in Heilongjiang Province, the S gene of a new strain NA33 was amplified, sequenced and analyzed. The results showed that the complete nucleotide sequence of the S gene of NA33 strain was composed of 1 693 nucleotides with TA-rich. The S gene contained one ORF, starting at position 37 and ending at position 1 326, encoding the N protein of 429 amino acid residues, and in line with HTN-based coding. Sequence comparison of the S genes between NA33 and reference hantavirus strains showed that NA33 was more homologous to Amur-like viruses than to the Hantaan (HTN) viruses or the other hantaviruses. Phylogenetic analysis of the amino acid sequence of N proteins showed that NA33 was clustered into the group of Amur-like viruses and was more similar to Far East Russia and Jilin strains isolated from Apodemus peninsulae. The phylogenetic tree indicated a certain degree of host-dependent characteristics and geographical aggregation characteristics of hantanviruses. Furthermore, the amino acid sequence of N protein of NA33 had the conserved amino acid sites of Amur-like viruses. In conclusion, Apodemus peninsulae carried Amur-like viruses in Heilongjiang province and was an important infectious source of hemorrhagic fever with renal syndrome (HFRS).
Subject(s)
Animals , Humans , China , Orthohantavirus , Chemistry , Classification , Genetics , Hantavirus Infections , Virology , Molecular Sequence Data , Murinae , Virology , Phylogeny , Rodent Diseases , Virology , Sequence Homology, Amino Acid , Viral Envelope Proteins , Chemistry , GeneticsABSTRACT
In order to determine the characteristics and genotypes of E protein genes of tick-borne encephalitis (TBE) virus strains DXAL-5, 12,13,16,18, 21 isolated from Ixodes persulcatus in the Northeast of China, cDNA synthesis of E protein genes of the six DXAL strains was performed using RT-PCR, and the E protein genes were cloned and sequenced. The results showed that the nucleotide sequence of E protein gene of the six DXAL strains was 1488 bp in length respectively and the length of predicted protein was 496 aa respectively. Sequence comparison of E protein genes among the six DXAL strains and the reference TBE virus strains showed that the six DXAL strains were more homologous to Far Eastern subtype strains than to Siberian subtype strains or European subtype strains. And the majority of subtype-determining amino acid sites of the six DXAL strains belonged to TBE virus Far Eastern subtype. Phylogenetic analysis of protein E showed that the six DXAL strains were all within the clade containing Far Eastern subtype strains. The new strains had higher identities and closer phylogenetic relationships with Senzhang strain, so we speculate that this vaccine strain still have good protection against the new TBE virus isolates. In the A, B and C antigenic domains of protein E, the six DXAL strains had different degrees of amino acid changes. These mutations were likely to affect the function of E protein.
Subject(s)
Animals , Mice , Amino Acid Sequence , Base Sequence , China , DNA, Complementary , Chemistry , Genetics , Encephalitis Viruses, Tick-Borne , Classification , Genetics , Molecular Sequence Data , Phylogeny , Reverse Transcriptase Polymerase Chain Reaction , Sequence Alignment , Sequence Analysis, DNA , Viral Envelope Proteins , Chemistry , GeneticsABSTRACT
192 samples of Masked Palm Civet (Paguma Larvata) from Guangdong Province were inoculated in Vero-E6 cells. One sample which came from masked palm Civet didn't cause cytopathic effects (CPE) until fourth-passage on Vero-E6 cells. Infected cells emerged granulating, shrinking, rounding and falling off. After three times freeze-thaw, cells and culture medium were harvested for electron microscopy. Virus particles were nonenveloped, double capsid and icosahedral symmetry. This virus was designated Masked Palm Civet/China/2004 (MPC/04). Hemagglutination test indicated that the virus could agglutinate healthy human type O red cells, but not the red cells of SPF chicken, experimental common bovine, rat and guinea pig. This virus was tolerant to chloroform treatment, pH 3.0 and water bath 50 degrees C 1 h. 1 M MgCl2 treatment could enhance resistance of virus to heat and increase infectivity. In order to classify the strain on the molecular level, specific primers according to mammalian reovirus were used for Reverse Transcription Polymerase Chain Reaction (RT-PCR). Appropriate specific products were amplified by RT-PCR. NCBI BLAST analysis indicated that this segment shared the highest identity to mammalian reovirus serotype 1 (T1L) virus. So we can deduce this virus is a member of the Reoviridae.