• Title/Summary/Keyword: Sequence-based molecular analysis

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Generation of Expressed Sequence Tags for Immune Gene Discovery and Marker Development in the Sea Squirt, Halocynthia roretzi

  • Kim, Young-Ok;Cho, Hyun-Kook;Park, Eun-Mi;Nam, Bo-Hye;Hur, Young-Baek;Lee, Sang-Jun;Cheong, Jae-Hun
    • Journal of Microbiology and Biotechnology
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    • v.18 no.9
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    • pp.1510-1517
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    • 2008
  • Expresssed sequence tag (EST) analysis was developed from three cDNA libraries constructed from cells of the digestive tract, gonad, and liver of sea squirt. Randomly selected cDNA clones were partially sequenced to generate a total of 922 ESTs, in which 687 unique ESTs were identified respectively. Results of BLASTX search showed that 612 ESTs (89%) have homology to genes of known function whereas 75 ESTs (11%) were unidentified or novel. Based on the major function of their encoded proteins, the identified clones were classified into ten broad categories. We also identified several kinds of immune-related genes as identifying novel genes. Sequence analysis of ESTs revealed the presence of microsatellite-containing genes that may be valuable for further gene mapping studies. The accumulation of a large number of identified cDNA clones is invaluable for the study of sea squirt genetics and developmental biology. Further studies using cDNA microarrays are needed to identify the differentially expressed transcripts after disease infection.

Analysis of Partial cDNA Sequence from Human Fetal Liver

  • Kim, Jae-Wha;Song, Jae-Chan;Lee, In-Ae;Lee, Young-Hee;Nam, Myoung-Soo;Hahn, Yoon-Soo;Chung, Jae-Hoon;Choe, In-Seong
    • BMB Reports
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    • v.28 no.5
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    • pp.402-407
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    • 1995
  • Single-run Partial cDNA sequencing was conducted on 1,592 randomly selected human fetal liver cDNA clones of Korean origin to isolate novel genes related to liver functions. Each partial cDNA sequence determined was analyzed by comparing it with the databases. GenBank, Protein Information Resource (PIR) and SWISS-PROT Protein Sequence Data Bank. From a set of 1.592 cDNA clones reported here, 1,433 (90.0% of the total) were informative cDNA sequences. The other 159 clones were identified as DNA sequences which had originated from the cloning vector. Among 1,433 informative partial cDNA sequences, 851 (59.3%) clones were revealed to be identical to known human genes. These known genes have been classified into 225 different kinds of genes. In addition, 340 clones (23.7%) showed various degrees of homology to previously known human genes. Ninety four (6.6%) clones contained various repeated sequences. Twenty four (1.7%) partial cDNA sequences were found to have considerable homology to known genes from evolutionarily distant organism such as yeast, rice, Arabidopsis, mouse and rat, based on database matches, whereas 124 (8.7%) had no Significant matches. Human homologues to functionally characterized genes from different organisms could be classified as candidates for novel human genes of similar functions. Information from the partial cDNA sequences in this study may facilitate the analysis of genes expressed in human fetal liver.

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Are the genus Moniliella and Trichosporonoides closely related in molecular taxonomic relationship\ulcorner (분자생물학적 방법을 이용한 Moniliella 속과 Trichosporonoides 속의 분류학적 고찰)

  • 신기선;신용국;태경환;권오유;이상한
    • Journal of Life Science
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    • v.11 no.2
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    • pp.133-137
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    • 2001
  • The molecular taxonomic relationship of nine species in the genus Moniliella Stolk & Dakin and Trichosporonoides Haskins & Spencer and six species of other yea나-like fungi was examined by sequencing analysis of large subunit rDNA D1/D2 variable domain. The fifteeen species fell into two major groups corresponding with their genetic relationships. The nine species of the genus Moniliella and Trichosporonoides were placed at the same cluster. similarity values based on the D1/D2 domain sequences were 45.4-100% among species of genus Moniliella, 45.2-84.4% among genus Trichosporonoides species, and 45.6-90.1% among species of genus Moniliella and Trichosporonoides. Identical sequence similarity was observed between M. suaveolens var. nigra and M. suaveolens. A colse relationship of M. mellis. and M. acetoabutens is observed. The result of this study provided and insight into the genetic origins of genus Moniliella and Trichosporonoides species as well as their genetic relationships. Genus Moniliella and Trichosporonoides are closely related to each other based on sequence analysis of the large subunit rDNA D1/D2 region and we suggest combination of the genus Moniliella and Trichosporonoides to single genus.

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Characteristics of Cucumber mosaic virus isolated from Zea mays in Korea

  • Kim, Mi-Kyeong;Kwak, Hae-Ryun;Lee, Su-Heon;Kim, Jeong-Soo;Kim, Kook-Hyung;Cha, Byeong-Jin;Choi, Hong-Soo
    • The Plant Pathology Journal
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    • v.27 no.4
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    • pp.372-377
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    • 2011
  • A virus causing mottle and stunt symptom on Zea mays was observed around Ulleng-do, Korea and identified as Cucumber mosaic virus (CMV-ZM) based upon biological, serological, and molecular characteristics. In host range studies, the CMV-ZM isolate produced local lesions on Datura stramonium, Vigna unguiculata, Cucurbita moschata, Chenopodium amaranticolor, Ch. quinoa, whereas this isolate produced systemic mosaic on Nicotiana tabacum cv. 'Xanthi-nc', Capsicum annuum, Solanum lycopersicum, Solanum melongena, Cucurbita pepo, and Z. mays. In addition, chlorotic local rings on inoculated leaves along with severe mosaic, malformation, and fern leaf symptoms on upper systemic leaves were shown in N. glutinosa plants. Complete nucleotide sequences of each genomic RNA segment was determined and compared to those of the other CMV strains. Comparison of the nucleotide sequence of 1a open reading frame (ORF) revealed approximately 89.2-92.4% sequence identity with each CMV subgroup IA and IB strain, while showing only 78% sequence identity with CMV subgroup II. Nucleotide sequence analysis of RNA2 ORFs revealed 85.3-97.6% sequence identity with subgroup I. In ORFs of RNA3, levels of nucleotide sequence identities were higher than 92-99.2% with CMV subgroup I and lower than 82% with CMV isolates of subgroup II. These results suggest that CMV-ZM isolate is more closely related to subgroup I than subgroup II and therefore, CMV-ZM isolate might be classified into as CMV subgroup I based on biological and molecular analysis.

Isolation of feline panleukopenia virus from Yanji of China and molecular epidemiology from 2021 to 2022

  • Haowen Xue;Chunyi Hu;Haoyuan Ma;Yanhao Song;Kunru Zhu;Jingfeng Fu;Biying Mu;Xu Gao
    • Journal of Veterinary Science
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    • v.24 no.2
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    • pp.29.1-29.12
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    • 2023
  • Background: Feline panleukopenia virus (FPV) is a widespread and highly infectious pathogen in cats with a high mortality rate. Although Yanji has a developed cat breeding industry, the variation of FPV locally is still unclear. Objectives: This study aimed to isolate and investigate the epidemiology of FPV in Yanji between 2021 and 2022. Methods: A strain of FPV was isolated from F81 cells. Cats suspected of FPV infection (n = 80) between 2021 and 2022 from Yanji were enrolled in this study. The capsid protein 2 (VP2) of FPV was amplified. It was cloned into the pMD-19T vector and transformed into a competent Escherichia coli strain. The positive colonies were analyzed via VP2 Sanger sequencing. A phylogenetic analysis based on a VP2 coding sequence was performed to identify the genetic relationships between the strains. Results: An FPV strain named YBYJ-1 was successfully isolated. The virus diameter was approximately 20-24 nm, 50% tissue culture infectious dose = 1 × 10-4.94/mL, which caused cytopathic effect in F81 cells. The epidemiological survey from 2021 to 2022 showed that 27 of the 80 samples were FPV-positive. Additionally, three strains positive for CPV-2c were unexpectedly found. Phylogenetic analysis showed that most of the 27 FPV strains belonged to the same group, and no mutations were found in the critical amino acids. Conclusions: A local FPV strain named YBYJ-1 was successfully isolated. There was no critical mutation in FPV in Yanji, but some cases with CPV-2c infected cats were identified.

Molecular Phylogenetic Study of Nesiohelix samarangae Based on CO-I Gene (동양달팽이 (Nesiohelix samarangae)의 CO-I 유전자를 이용한 분자계통학적 연구)

  • Bang, In Seok;Lee, Yong Seok
    • The Korean Journal of Malacology
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    • v.30 no.4
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    • pp.391-397
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    • 2014
  • Previously, we have reported expressed sequence tags (ESTs) analysis on the land snail, Nesiohelix samarangae (Ns). Of these ESTs, we have identified four partial fragments of N. samarangae cytochrome oxydase I (NsCO-I) gene which lead to obtain an 852 bp partial cDNA. Since NsCO-I is one of the best-known molecular phylogenetic markers, we have attempted to conduct comparative in silico analysis by using the NsCO-I gene. The combined results from BLAST analyses, multiple sequence alignment and molecular phylogenetic study of NsCO-I cDNA indicate that N. samarangae has similarity to three land snails such as Elona quimperiana, Euhadra herklotsi and Euhadra idzumonis.

Molecular Phylogeny of Silk Producing Insects Based on Internal Transcribed Spacer DNA1

  • Mahendran, Botlagunta;Ghosh, Sudip K.;Kundu, Subhas C.
    • BMB Reports
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    • v.39 no.5
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    • pp.522-529
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    • 2006
  • Silk moths are the best studied silk secreting insects and belong to the families Bombycidae and Saturniidae. The phylogenetic relationship between eleven silk producing insects was analyzed using the complete DNA sequence of the internal transcribed spacer DNA 1 locus. The PCR amplification and sequence analysis showed variation in length ranging from 138 bp (Antheraea polyphemus) to 911 bp (Hyalopora cecropia). Microsatellite sequences were found and was be used to distinguish Saturniidae and Bombycidae members. The nucleotide sequences were aligned manually and used for construction of phylogenetic trees based on Maximum parsimony and Maximum likelihood methods. The topology in both the approaches yielded a similar tree that supports the ancestral position of the Antheraea assama.

Analysis of the Complete Genome Sequence of Zucchini yellow mosaic virus strain A Isolated from Hollyhock

  • Choi, Seung-Kook;Yoon, Ju-Yeon;Sohn, Seong-Han
    • The Plant Pathology Journal
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    • v.23 no.4
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    • pp.245-250
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    • 2007
  • The complete genome sequence of Zucchini yellow mosaic virus stain A (ZYMV-A) isolated from a hollyhock (Althaea rosea) was determined by using RT-PCR with a series of primer sets. The virus genome consisted of 9593 nucleotides (nt), excluding the poly(A) tract at 3' terminus of the virus genome, with 5' and 3' untranslated region of 139 and 211 nt, respectively. The deduced polyprotein of ZYMV-A consisted of 3080 amino acid (aa) residues and was 351 kDa in molecular weight. All proteolytic cleavage sites of the polyprotein of ZYMV-A were compared with those of ZYMV strains, which showed the cleavage sites were conserved among ZYMV strains. The HC-Pro contained the KITC and PTK motifs, and the DAG motif was located at CP ORF of ZYMV-A, suggesting that ZYMV-A is aphid-transmissible. Phylogenetic tree analysis based on the complete genome among ZYMV strains or CP ORFs with other potyviruses showed ZYMV strains formed a distinct group. These results clearly confirmed that ZYMV-A was another distinct strain in ZYMV population at molecular level.

Non-pungent Capsicum Contains a Deletion in the Capsaicinoid Synthetase Gene, which Allows Early Detection of Pungency with SCAR Markers

  • Lee, Choong-Jae;Yoo, Eun Young;Shin, Joo Hyun;Lee, Jemin;Hwang, Hee-Sook;Kim, Byung-Dong
    • Molecules and Cells
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    • v.19 no.2
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    • pp.262-267
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    • 2005
  • The capsaicinoid synthetase (CS) gene cosegregated perfectly with the C locus, which controls the presence of pungency, in 121 $F_2$ individuals from a cross between 'ECW123R' and 'CM334', both of Capsicum annuum. We concluded that CS and C are tightly linked. Sequence analysis of the genes of four pungent and four non-pungent pepper lines showed that the non-pungent peppers had a 2,529 bp-deletion in the 5' upstream region of CS. We have developed molecular markers of the C locus to detect pungency at the seedling stage. Based on the deleted sequence, we developed five SCAR markers, two of them being codominant. These SCAR markers will be useful for easy, accurate, and early detection of non-pungent individuals in breeding programs.

Gene Identification and Molecular Characterization of Solvent Stable Protease from A Moderately Haloalkaliphilic Bacterium, Geomicrobium sp. EMB2

  • Karan, Ram;Singh, Raj Kumar Mohan;Kapoor, Sanjay;Khare, S.K.
    • Journal of Microbiology and Biotechnology
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    • v.21 no.2
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    • pp.129-135
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    • 2011
  • Cloning and characterization of the gene encoding a solvent-tolerant protease from the haloalkaliphilic bacterium Geomicrobium sp. EMB2 are described. Primers designed based on the N-terminal amino acid sequence of the purified EMB2 protease helped in the amplification of a 1,505-bp open reading frame that had a coding potential of a 42.7-kDa polypeptide. The deduced EMB2 protein contained a 35.4-kDa mature protein of 311 residues, with a high proportion of acidic amino acid residues. Phylogenetic analysis placed the EMB2 gene close to a known serine protease from Bacillus clausii KSM-K16. Primary sequence analysis indicated a hydrophobic inclination of the protein; and the 3D structure modeling elucidated a relatively higher percentage of small (glycine, alanine, and valine) and borderline (serine and threonine) hydrophobic residues on its surface. The structure analysis also highlighted enrichment of acidic residues at the cost of basic residues. The study indicated that solvent and salt stabilities in Geomicrobium sp. protease may be accorded to different structural features; that is, the presence of a number of small hydrophobic amino acid residues on the surface and a higher content of acidic amino acid residues, respectively.