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PubMed2005

[Identification and characterization of a novel gene EOLA1 stimulating ECV304 cell proliferation].

OBJECTIVE: To amplify the full-length cDNA and characterize the structure and biological function of a novel expression sequence tag ST55 (GenBank Accession No. BM121646). METHODS: Rapid amplification of cDNA ends was used to clone the full-length of cDNA of ST55 in this study. Then, its tissue distribution was checked by Northern blots, and the associated protein was screened by GAL 4-based yeast two-hybrid. The effect of stable transfection of the cDNA on cell proliferation was evaluated in ECV304 cells. RESULTS: A full-length 1404 bp cDNA was cloned, and it was accepted as a novel human mRNA by GenBank (No. AY074889), named endothelial-overexpressed lipopolysaccharide-associated factor 1 (EOLA1). Bioinformatic analysis found that the EOLA1 encoded 158 amino acids, 17.89 kDa protein, and mapped to chromosome Xq27.4 with 5 exons. EOLA1 expressed in different human normal tissues and cancer cell lines. Using the EOLA1 cDNA as bait, we performed a yeast two-hybrid screening of a human liver cDNA library and identified metallothionein 2A (MT2A) as associated protein. The interaction between EOLA1 and MT2A was confirmed by co-immunoprecipitation experiments. Stable transfection of EOLA1 was noted to stimulate ECV304 cell proliferation (P < 0.05). CONCLUSION: The findings suggest that EOLA1 is a novel gene and the interaction of EOLA1 and MT2A may play an important role in cell protection in inflammation reaction.

Amino Acid Sequence↗
PubMed2005

[Cloning and bioinformatic analysis of a novel gene HCR2 up-regulated in aorta of hypercoagulable rat].

OBJECTIVE: The aim of this study was to clone the full-length cDNA of HCR2 up-regulated in aorta of hypercoagulable rat and make the relevant bioinformatic analysis. METHODS: Rapid amplification of cDNA end (RACE) and nested PCR technique was used to amplify the full-length cDNA of HCR2 from an EST (GenBank accession number BQ901227) which was significantly up-regulated in hypercoagulable rat. RESULTS: The full-length cDNA of HCR2 has been obtained (GenBank accession number AY234417). Semi-quantitative RT-PCR demonstrated that HCR2 is up-regulated in aorta of hypercoagulable rat. Bioinformatic analysis showed that HCR2 gene is located in rat chromosome 4q11. The full-length cDNA of HCR2 is 1275 bp, coding for a 78 amino acids polypeptide with a theoretical molecular weight of 8841.7 and isoelectric point of 8. 59. This protein has no signal peptide and transmembrane sequence. It may be a nuclear protein. It shares no significant homology with any known protein. CONCLUSION: The successful cloning of HCR2 has laid a solid foundation for further study of its function and its possible role in the occurrence and development of hypercoagulability.

Amino Acid Sequence↗
PubMed2005

[Cloning and expression in Pichia pastoris of an alkaline mannanase gene].

A strain containing alkaline mannanase gene was isolated from soil by functional plates and the genome library was constructed. From it a mannanase gene TM1 was acquired and was sequenced. The BLAST analysis showed a lower-than-60% similarity of the amino acid sequence to those in GenBank and proved TM1 to be a new mannanase gene (GenBank accession number AY623903). The new gene without signal peptide was cloned into the Pichia pastoris expression vector pHBM905C. The recombinant plasmid pHBM1201 was digested by Sal I and transformed into Pichia pastoris KM71, GS115, SMD1168, respectively. All of the recombinant Pichia pastroris strains containing pHBM1201 secreted functional beta-mannanase. Because of its high mass of expression, the recombinant Pichia pastoris SMD1168-3 containing pHBM1201 was induced at shake flasks. The optimal temperature and pH of the beta-mannanase produced by the recombinant strains were 55 degrees C and 7.5, respectively. The enzymatic activity for konjak powder reached 41.8 with a half life of one hour. After keeping at 80 degrees C for 5 min, the enzymatic activity declined from 77% to 11% and the enzymatic activity could recover up to more than 60% when the temperature descended to 55 degrees C.

Cloning, Molecular↗
PubMed2005

[Expression of aquaporin 2, a water channel, in human endometrium during the menstrual cycle].

OBJECTIVE: To examine the expression of aquaporin 2 (AQP2) in human endometrium. METHODS: Specimens of human endometrium were collected from 87 women at different menstrual cycles, aged 30 +/- 3, 23 cases in the proliferative phase, 30 cases in the early secretory phase and 34 cases in the mid-secretory phase respectively. Immunohistochemistry and Western blotting were utilized to detect the localization and expression of AQP2 in the endometrium. Reverse transcription-polymerase chain reaction (RT-PCR) was employed to detect its messenger RNA. The PCR product was cloned and sequenced. RESULTS: Positive immunoreactivity of AQP2 was found in the epithelia cells and glandular epithelial cells of all specimens of human endometrium at different phases of the menstrual cycle, and all stromal cells were not stained. The reactive substance was primarily distributed in the membrane and cytoplasm, but not in the nuclei of all positive cells. In Western blotting showed dominant bands with relative molecular weight between 35,000 Da and 50,000 Da, which was corresponded to the glycosylated form of AQP2 by the positive control from rat kidney. Semi-quantitative analysis showed that the relative expression of AQP2 in the mid-secretory phase was 1.63 +/- 0.15, significantly higher than those in the early secretory phase (1.33 +/- 0.14, P < 0.05) and that in the proliferative phase (1.03 +/- 0.10, P < 0.01). Message RNA was found out in all cases by RT-PCR and the PCR product was confirmed in nearly exact (99%) consistency with the GenBank by sequencing. AQP2 mRNA was expressed in all normal endometrium at different phases, and was weakly expressed in the endometrium at the proliferative phase. Sequencing showed that the AQP2 sequence was 99% homologous with that in the GenBank. CONCLUSION: AQP2 expression in the human endometrium suggests that AQP2 may be involved in the regulation of uterine fluid homeostasis influenced by ovarian steroid hormones in the menstrual cycle.

Adult↗
PubMed2005

[Screening and identification of human nasopharyngeal carcinoma-associated antigens].

OBJECTIVE: The purpose of this study was to provide an informative marker for clinical immunopathological diagnosis of nasopharyngeal carcinoma (NPC) and to lay a foundation for furture study of immunotherapy. METHODS: The NPC cDNA expression library was screened by serological analysis of recombinant cDNA expression libraries (SEREX) technology using xenogenic immune sera. The positive nucleotide sequences of cDNA inserts were determined and analysed with DNASIS and BLAST software in EMBL and GenBank. RESULTS: Eighteen positive clones encoding antigen genes were obtained after immunoscreening. These antigen genes included known genes, such as MAGE gene, CREM gene, beta2-microglobulin, Ribosomal Protein, et al, and unknown genes or ESTs in GenBank. To characterize the expression profile of these genes, the antibodies in sera from 40 nasopharyngeal carcinoma patients and 40 healthy donors were assayed with three antigens to screen specific and relative serum markers for nasopharyngeal carcinoma. The results showed that the positive rates for These results indicate nasopharyngeal carcinoma patients were higher than those for healthy donors. CONCLUSION: that some of the antigens may be related to nasopharyngeal carcinoma and may be valuable tumor markers in the diagnosis of nasopharyngeal carcinoma.

Antigens, Neoplasm↗
PubMed2006

[Screening and preliminary analysis of colorectal carcinoma-associated antigen genes].

OBJECTIVE: To screen and identify the genes coding for colorectal carcinoma-associated antigen and analyze the bioinformation of their cDNA sequences. METHODS: Immunoscreening of the cDNA phage-display library derived from human colorectal carcinoma was performed with autologous or allogeneic serum antibody from patients with colorectal cancer through SEREX approach. After amplification of the positive phage clones, the phage DNA was extracted and purified with Qiagen kit, and the fragment sizes of the cDNA of positive clones were identified by PCR and EcoR I and Hind III restriction endonucleases. The cDNAs of the positive clones were ligated into pUCm-T vector and sequenced. The bioinformation of cDNA sequences were analyzed against GenBank+EMBL+DDBJ+PDB Sequences Database. RESULTS AND CONCLUSION: Eleven positive clones were obtained after immunoscreening, and the sizes of the cDNA fragments were 1100, 1300, 1000, 2000, 1200, 1200, 700, 900, 600, 1200 and 1000 bp, respectively, representing 9 antigen genes, including 7 with homology with the known genes. Among the 11 obtained positive clones, 3 were the same cDNA having homology with interferon-induced transmembrance protein-1 and possessing anti-proliferation effect; another 6 represented different genes, namely human BAC clone RP11-453E17 whose function have not been cleared, human cartilage-hair hypoplasia region gene responsible for cartilage-hair hypoplasia, human chromosome 5 clone CTD-2030B15 with insertion mutation, human gene similar to anti tumor necrosis factor-alpha antibody light-chain Fab fragment associated with tumor growth, mRNA of human beta-2-microglobulin in relation to tumor cell proliferation, and human aldolase A gene promoting tumor cell proliferation. The other two cDNA sequences were not identified for homology with currently known genes in GenBank, and their functions awaits further investigation.

Antigens, Neoplasm↗
PubMed2006

Genetic analysis of CAV1 gene in hypertension and metabolic syndrome.

Recently, we reported that the polymorphism 1132T>C (GenBank: AF519768.1) of the NOS3 gene was associated with susceptibility to metabolic syndrome (MS) in hypertensive patients. This suggests that other genes such as CAV1, whose product (CAV1) regulates eNOS activity, could also be related to this phenotype. In this work we investigated the following: i) whether CAV1 is a quantitative trait locus of clustering of atherothrombotic traits associated with MS; ii) whether CVA1 is associated with hypertension or MS in hypertensive patients; and iii) whether genetic interaction between NOS3 and CAV1 is involved in the susceptibility or protection to hypertension associated with MS. To carry out the study, we genotyped 285 randomly selected individuals and 175 hypertensive patients, all of them < or = 60 years old, with two polymorphisms of the CAV1 gene: the 22285 C>T and the 22375-22375 del AC (GenBank AF125348), and the 1132T>C polymorphism of the NOS3 gene. To perform sample genotyping, we used pyrosequencing and FRET techniques. The 22285 C-22375-22375 del (Cd) haplotype of CAV1 gene was associated with low levels of blood pressure in the general population. Moreover, it was a genetic protection factor against MS in hypertensive patients. In addition, we found no evidence of gene-gene interaction between NOS3 and CAV1 genes with regard to that phenotype.

Case-Control Studies↗
PubMed2006

[Cloning and sequence analysis of a novel member of the rab gene family from Euplotes octocarinatus].

Rab proteins belong to a subfamily of small GTP-binding proteins of the Ras superfamily, which play an important role in intracellular vesicular traffic. In this study, a rab gene was obtained from Euplotes octocarinatus by polymerase chain reaction (PCR) and RT-PCR. The rab gene from macronucleic DNA was 884 bp in length, including non-coding regions and telomeric sequences at both ends. The rab gene from micronuclear DNA (723 bp), lacking of internal eliminated sequences, was identical to rab gene from macronuclear DNA. RT-PCR showed that the opening reading frame of the rab gene was 663 bp long. The rab gene from macronuclear DNA contained an intron of 60 bp at the position from 153 bp to 212 bp of macronuclear DNA. The rab gene had two in-frame TGAs encoding for cysteine in Euplotes octocarinatus. The rab gene used TAG as stop codon, which was the first report in Euplotes octocarinatus. The result of BLAST in NCBI demonstrates that the Rab shares a homology of 49-52% at the amino acid level with Rab1 proteins from a number of other eukaryote, which suggesting that the Rab is a Rab1 homolog. The rab gene was therefore designated Eo-rab-1N (GenBank accession number: DQ105562). The evolution of Eo-rab-1N was analyzed using phylogenetic tree of amino acids sequences of Rab1 obtained from GenBank.

AT Rich Sequence↗
PubMed2006

[Molecular cloning and expression analysis of a SUPERMAN-like zinc finger protein gene in upland cotton].

The zinc finger proteins belong to the largest family of regulatory transcription factors, which play an important role in growth and development in animal and plant systems. SUPERMAN-like zinc finger protein gene has only one "finger like" motif. A pair of degenerate primers was designed according to the conserved regions, and 3 kinds of EST of this family were isolated from cotton through RT-PCR. The full length of one SUPERMAN-like zinc finger protein also has been acquired. The entire coding region is 744 bp and encodes a polypeptide of 248 amino acids with 40% homology to RBE protein of Arabidopsis deposited in the GenBank. This gene was designated as GZFP. It has the conserved zinc finger domain and the leucine rich region at the carboxyl terminus but no intron in the coding region. GZFP also has the plant nuclear localization signal. GZFP shows a more expression pattern in floral buds, ovaries, petals and roots than in phloem, xylem, fibers, leaves and seeds of cotton by RT-PCR, although it has a very low detection level and there is not any homologous ESTs found in the GenBank. Analysis of the 5' flanking sequence shows there are several regulatory elements responsible for pollen and root expression, four core sites required for binding of Dof proteins and four light-regulated elements.

5' Flanking Region↗
PubMed2005

Identification and characterization of Bombyx mori eIF5A gene through bioinformatics approaches.

As the genome of B. mori is available in GenBank and the EST database of B. mori is expanding, identification of novel genes of B. mori was conceivable by data-mining techniques and bioinformatics tools. In this study, we used the in silico cloning method to identify eukaryotic initiation factor 5A (eIF5A) gene in B. mori. With the hypusine formation, eIF5A is involved in the regulation of cell proliferation and apoptosis. Using the computer program MEGA3, we conducted a search for homologs of eIF5A among many eukaryotic species and confirmed that the eIF5A was conserved in all organisms investigated. This gene has been registered in GenBank under the accession number DQ104412.

Amino Acid Sequence↗
PubMed2006

[Human coronavirus-NL63 was detected in specimens from children with acute respiratory infection in Beijing, China].

OBJECTIVE: A new human coronavirus, HCoV-NL63, was identified recently from two Dutch children with acute respiratory infection (ARI) by two scientists in the Netherlands in 2004. To investigate if this newly discovered virus is associated with acute respiratory infections in pediatric patients in Beijing, tests were developed to detect HCoV-NL63 gene fragments from throat swab and nasopharyngeal aspirates collected from children in outpatient and inpatient departments with ARI in Beijing from Dec. 2003 to Mar. 2004. METHODS: A total of 245 clinical samples, which were negative either for diagnostic tests of human respiratory syncytial virus, influenza virus A and B, adenovirus, parainfluenza virus 1, 2 and 3 by indirect immunofluorescence assay or human metapneumovirus by RT-PCR, were screened for HCoV-NL63 by nested PCR amplifying gene fragments located on the 1b and 1a genes. Amplicon of PCR from 1a gene of HCoV-NL63 was sequenced and the sequences were compared with those in GenBank nucleotide sequence database. RESULTS: Three (1.2%) out of the 245 samples were positive for HCoV-NL63 by nested-PCR using primers on 1b gene. These three samples also showed positive results on nested PCR in which primers were designed with sequences complementary to 1a gene segments. These positive samples were collected from hospitalized children under 2 years of age with pneumonia, bronchiolitis and bronchitis, respectively. The partial 1a gene sequences from two positive samples (BJ3140 and BJ3787) of HCoV-NL63 showed 100% homology between each other and high homology (98%-99%) with the sequences of 1a gene of HCoV-NL63 reported from different countries in GenBank. Phylogenetic analysis showed that BJ3140 and BJ3787 fell into the same genetic cluster (group 1). CONCLUSIONS: These data suggest that some of acute respiratory infections in young children in Beijing area are related to the newly identified HCoV-NL63.

Acute Disease↗
PubMedDate not supplied

A novel polymorphism of the porcine prolactin gene (PRL).

Prolactin is a protein hormone playing a role in the maintenance of pregnancy in the pig by action on corpora lutea cells and possibly initiating production of progesterone. The prolactin gene is 10 kb in size and is composed of 5 exons and 4 introns. The present work is a report of the swine PRL gene--comparative DNA sequence analysis and the SNP revealed in the promoter region. Based on the bovine prolactin gene, three primer pairs were designed using the Primer3 on-line software. The overlapping fragments covered about 400 nucleotides of the promoter and 78 nucleotides of exon 1. The fragments were amplified; two of them were sequenced and deposited in the GenBank database (AY341908 and AY905690). All fragments were analyzed using multitemperature SSCP (MSSCP) technique. Only one fragment appeared to show a different MSSCP pattern. The samples of differing MSSCP conformers were sequenced and the C499T transition was identified in the 5'UTR region of the gene. The HphI restriction enzyme appeared to recognize the novel SNP. The alignment for homology analysis was performed with porcine, bovine (X01452) and human (NM_000948) DNA sequences available in GenBank database, using BLAST software. The comparative homology analysis results varied in dependence on the species and functional region of the gene.

5' Untranslated Regions↗
PubMed2006

[Identification of hepatitis B virus genotypes in patients with chronic hepatitis B from different nationalities in ethnic minority areas in Yunnan Province, China].

OBJECTIVE: To determine whether there is evolutionary difference in hepatitis B virus (HBV) genotypes among the patients with chronic hepatitis B (CHB) of different nationalities and its clinical significance. METHODS: Peripheral blood samples were collected from 50 CHB patients, 25 of diverse nationalities and 25 of Han nationality from the ethnic minority regions in Yunnan Province, China, The HBV preS2/S (pre S2/S) and C genes were amplified by PCR. The PCR products were inserted into the vector pBluescriptIISK (pBS). The cloned preS2/S and C genes were sequenced. RESULTS: The sequences of HBV preS2/S and C genes from the 50 patients were 846 (with 49.1% of GC) and 552 (with 46.1% of GC) nucleotides (nt) in length, and encoded 281 and 183 amino acids (aa) respectively. These findings were registered in GenBank Accession Numbers: AY517619, AY517620, AY517488, AY517489, AY517598, AY517599. Compared with the HBV and subtype sequences in the GenBank database, the HBV preS2/S and C genes among all the subjects were homologous to ayw1 in sequence by 97.5% - 98.6% and 94.5% - 97.8% respectively. The "a" determinant region of S genes in all cases were found to be Arginine (AGA) and Lysine (AAA) at corresponding aa 122nd and 160th respectively. HBV genotype B was identified in all patients with CHB (ayw1 subtype). Genotypes A, C, D, E, F, G, and H were not detected in any of them. The quasi-species nature of the HBV in the sera was observed in 2 of the 50 samples examined (4%). There was not a significant difference in the prevalence of HBV genotype B between the 25 diverse nationality patients and the 25 control Han nationality patients (P > 0.05). In the 50 CHB patients, the preS2/S genes were identified to have aa substitutions at the positions R124K (1.1%), L172P (1.3%), M306T (1.5%), and I361M (1.6%), with a frequency of more than 1%. In all subjects, the frequency of aa G145R (0.4%) substitutions was less than 1%. In all subjects, nt variations of C genes caused aa substitutions among aa 27 - 63, 80 - 110, and 135 - 153 involved in T and B cell epitopes. In 45 CHB patients, C genes was identified to have aa substitutions at the positions V 27, N38, V63, Q135, and A147, due to nt variations of 1979A to G, 2012T to A, 2088G to T, 2304C to A, and 2339A to G transitions respectively. The frequency of aa substitutions of C genes was more than 1%. Whereas as for the other 5 severe CHB patients, the C gene variations of A to G, and A to C transitions at nt positions 2159 and 2189 led to aa substitutions of S to G and I to L at positions G87 and L97. No insertion or deletion was found in preS2/S and C regions. HBV genotype B was not relevant to different nationalities (all P > 0.05). CONCLUSION: It is the first time that the genotype of the HBV epidemic strains in the ethnic minority areas of Yunnan Province has been identified as genotype B subtype ayw1. The HBV genotype B is not related with nationality. A novel genotyping method by using PCR, gene cloning, followed by DNA sequencing that can identify all major genotypes has been developed. HBV genotype B is the geographic original strain in this area and is correlated with the severity of liver diseases and curative effect. HBV viral is the only significant variable associated with the CHB patients' prognosis.

Adolescent↗
PubMed2006

[DNA sequence polymorphism of Chlamydia trachomatis omp1 gene].

A method for detection and genotyping of genital Chlamydia trachomatis infections based on omp1 gene amplification and sequencing was developed, and the character of omp1 gene of Chlamydia trachomatis was analysed. Urethral or endocervical specimens were collected from 323 patients attending STD clinics in Hengyang, Shanghai, Guangzhou and Jiangmen from November, 2003 to May, 2004. DNA was extracted by usual method, and an approximately 980bp fragment from the major outer membrane protein (omp1) gene of Chlamydia trachomatis was amplified by nested polymerase chain reaction (nPCR). The PCR products were purified by DNA agarose gel purification system and the sequence of the omp1 gene was determined by using an ABI PRISM 3700 Genetic Analyser, and genotyping was performed by BLAST similarity search. Multiple alignment was performed with CLUSTAL package (CLUSTAL X), and a phylogenetic tree was constructed by Mega3 software to illustrate the evolutionary relationships between clinical isolates and reference strains of C. trachomatis obtained from GenBank. All variable sequences were submitted to GenBank by Banklt programe. The overall prevalence of urogenital chlamydial infection was 29.7% (96 of 323). All the 96 C. trachomatis-positive cases were sequenced, and 10 genotypes and 28 genetic variants were detected. The most prevalent genotype was E(34.4%), followed by J(25.0%), D(12.5%), F(8.3%), G(7.3%), H(3.1%), Ba(3.1%), K(3.1%), Da (2.1% ), 1 (1.1%). The distribution of C. trachomatis genotypes in the four cities in sourth China was similar to other countries in the world. The omp1 gene was highly conserved for genotype E and F, but appeared slightly less conserved for other genotypes, where the sequences displayed one or several nucleotide substitutions relatived to the corresponding reference sequence. And a similar recombination was found between genotypes Ba and D in CD1. Phylogenetic tree showed that Chlamydia trachomatis genotypes were mainly divided into three clusters, according to previous grouping in the B, F-G, and C complexes. Clusters F-G and C were characterized by small genetic distances within each cluster, but clusters B displayed larger genetic distances. And the clinical isolates were highly related to the reference strains. It is concluded that the isolated Chlamydia trachomatis strains exhibit remarkable omp1 DNA sequence polymorphism, which can encourage for vaccine design and infection control.

Chlamydia Infections↗
PubMed2006

[Investigation on hantaviruses infection in rodents from free markets in Beijing areas].

OBJECTIVE: In order to find out the factors related to hemorrhagic fever with renal syndrome (HFRS) infection, and to evaluate the probability of ecdemic hantaviruses (HV) infection in rodents in Beijing areas. METHODS: Rodents were collected in a large-scale railway station and a produce market with 'trap nights' method from April to May, 2004. The IgG reacting sera to HV antigen were detected using ELISA. The partial M and S segment of HV from captured rodent lung samples were amplified with RT-PCR. The PCR products were purified and sequenced. BLAST program was then used to perform on nucleotide pairwise alignment with all available sequence in GenBank. The alignment of the multiply nucleotide and the deduced amino acid sequences, together with phylogenetic analysis were completed with DNASTAR software. RESULTS: The average population density was 3.49% (24/690). The overall seroprevalence of HV infection was 8.3% (2/24). RT-PCR positive rates were 8.3% (2/24). The nucleotide sequences of 356 bp region (1958 - 2313) of M segment obtained from 2 samples were all identified to Seoul virus (SEOV), with 7.6% heterogeneity. The dc501 strain from railway station was closely related to SD227 and Hebei4 from Shandong and Hebei provinces respectively. BjFT01 strain from the farm product market had more special nucleotide transitional mutations than other known SEOV from Beijing in GenBank. This strain, together with known HN71 from Hainan province, K24-E7 from Zhejiang province, L99 from Jiangxi province and R22 from Henan province, represented a monophylogentic linkage. CONCLUSION: The higher HV prevalence of rodents in transportation center was the potential and important risk for HFRS epidemic in Beijing. The increasing prevalence of M. musculus should call for attention. It was possible that SEOV in Beijing was imported by infected rodents through vehicles from other provinces.

Animals↗
PubMed2006

[Sequence and structure analysis of mitochondrial tRNApro and tRNAthr genes in domestic goose breeds].

We report here the results of the sequence and structure analysis of mitochondrial tRNApro and tRNAthr genes in domestic goose breeds by sequencing the mitochondrial DNA from a total of 25 samples from 6 breeds of Chinese geese and 2 breeds of domestic Europe geese. Sequences and the cloverleaf structure of tRNApro (69 bp) and tRNAthr (68 bp) in domestic goose breeds were described and analysed They were compared amongst the three domestic goose breeds as well as between Anseriformes (Anser cygnoides) and Galliformes (Gallus gallus domesticus, Genbank accession number NC001323). Both goose tRNApro and tRNAthr genes have normal cloverleaf secondary structures. The amino acid arm and the anticodon loop of the cloverleaf structure of tRNApro and tRNAthr are very conservative among Anser albifrons, Anser anser and Anser cygnoides. The gene sequences in this study were deposited to GenBank under accession numbers AY427800-AY427805 and AY427812-AY427814.

Animals↗
PubMed2006

[Effects of MTNR1A gene on litter size in a large white and a landrace herd].

Two pairs of primers were designed based on the known sequence in GenBank for amplification of MTNR1A gene in a Large White and a Landrace herd. Using PCR-SSCP (single strand conformation polymorphism), we found a single nucleotide polymorphism(SNP) within the product amplified from the first pair of primers. PCR products from randomly selected different genotypes were sequenced after were recovered and purified. Results revealed a synonymous single base mutation(G-->A) at +159bp(sequence numbering based on Genbank accession number U73326) for the BB genotype. When analyzed for association with litter size traits, this MTNR1A SNP was found to have no significant effect on litter size traits.

Animals↗
PubMed2006

[Mixed infection of Echinococcus granulosus and Echinococcus multilocularis in dog].

OBJECTIVE: To identify mixed infection of Echinococcus granulosus and E. multilocularis in a dog from Xinjiang. METHODS: Thirty dogs from the pasture area were dissected and over 10,000 Echinococcus adult worms were found from one dog. Morphological observation revealed possible mixed infection of the two Echinococcus species. Further identification was made by amplification of the target gene DNA fragment (mitochondrial 12S rRNA gene). RESULTS: The adult worms of E. granulosus showed a relatively longer and larger gravid proglottid, its genital pore situated near or below the middle-side of the segment. The uterus was in a sacculate shape with irregular branches and approximately over 200 - 800 eggs in it. Morphology of the adult worms of E. multilocularis was similar to E. granulosus, slightly smaller, consisting of 4 to 5 proglottids. The uterus was not sacculate and with no branch. Its lateral genital pore often situated in the anterior part of the segment. Sequence analysis of mitochondrial 12S rRNA gene showed that amplification with the Eg1f/r primers shared complete identity with E. granulosus G1 genotype (GenBank accession no. AY462129), while that witht the EmH15/17 primers shared complete identity with E. multilocularis (GenBank accession no. AB031351). The presence of both E. granulosus and E. multilocularis was confirmed by microscopy and gene identification. CONCLUSION: Mixed infection of the two species of Echinococcus has been confirmed in the dog by morphological observation and PCR technique.

Animals↗