Molecular cloning and characterization of an antigenic protein with a repeating region from Clonorchis sinensis

Kim, Tae Yun; Kang, Sung-Soo; Ahn, Il Young; Cho, Seung Yull; Hong, Sung-Jong

doi:10.3347/kjp.2001.39.1.57

Cited by 18 publications

(16 citation statements)

References 32 publications

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“…It was also found that the ESTs contained several antigenic protein genes for C. sinensis or other parasites. The recombinant glycine-rich C. sinensis protein and proline-rich antigen were reported previously to be useful for immunodiagnosis, with a high specificity (Yang et al 2000;Kim et al 2001).…”

Section: Discussionmentioning

confidence: 98%

Analysis of the genes expressed in Clonorchis sinensis adults using the expressed sequence tag approach

Lee

Park

et al. 2003

Parasitol Res

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Clonorchis sinensis is a biliary tract parasite, which infects over 30 million people in China, Korea and Southeast Asia through the ingestion of undercooked freshwater fish that harbour the infective metacercariae. The genes expressed in C. sinensis adults were identified in order to develop novel drugs, better diagnostics and vaccines for the parasite. The C. sinensis cDNA library was constructed and DNA sequencing was performed with 450 randomly selected clones. Four hundred and fifteen clones contained the amino-acid-encoding sequences. The functions of these genes could be assigned by DNA sequence homology. Basic Local Alignment Search Tool X analysis showed that 277 of the 415 clones were strongly matched ( P<10(-9)) to previously identified proteins, while the remaining 138 fell into the "no database match" category. Among the clones matching previously identified proteins, 220 putatively identified genes were sorted into seven functional categories. These included the genes associated with energy metabolism (38), gene expression/RNA metabolism (21), regulatory/signalling components (14), protein metabolism/sorting (98), the structure/cytoskeleton (29), membrane transporters (ten) and antigenic proteins (ten). The remaining 57 clones were not included in these categories. The dataset included the genes encoding the proteases, a lipid binding protein, the antigen proteins and the other genes of interest from a diagnostics, drug or vaccine development viewpoint. The present expressed sequence tag analysis proved to be an effective tool for examining gene expression and identified several important genes which increase and complement our knowledge of the biology of C. sinensis.

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Section: Discussionmentioning

confidence: 98%

Analysis of the genes expressed in Clonorchis sinensis adults using the expressed sequence tag approach

Lee

Park

et al. 2003

Parasitol Res

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“…Furthermore, recombinant GRCsP protein showed high sensitivity and specificity by ELISA for clonorchiasis [87,88]. Furthermore, a proline-reach antigen (CsRPA) containing 15 GPDAPVPKSG repeats was found to have high sensitivity and specificity against clonorchiasis sera [89].…”

Section: Excretory-secretory (Es) Antigensmentioning

confidence: 96%

Functional Genes and Proteins of Clonorchis sinensis

Kim

Hong

2009

Korean J Parasitol

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During the past several decades, researches on parasite genetics have progressed from biochemical and serodiagnostic studies to protein chemistry, molecular biology, and functional gene studies. Nowadays, bioinformatics, genomics, and proteomics approaches are being applied by Korean parasitology researchers. As for Clonorchis sinensis, investigations have been carried out to identify its functional genes using forward and reverse genetic approaches and to characterize the biochemical and biological properties of its gene products. The authors review the proteins of cloned genes, which include antigenic proteins, physiologic and metabolic enzymes, and the gene expression profile of Clonorchis sinensis. CHROMOSOMES AND GEOGRAPHICAL ISOLATESC. sinensis has 2n = 56 chromosomes, where n consists of 8 large and 20 small chromosomes. C. sinensis geographical isolates collected in Korea and northeastern China (Liaoning Province) have all demonstrated the same chromosome number, but different karyotypes. Specifically, Korean isolates have 3 metacentric and 1 meta-/submetacentric pairs, whereas Chinese isolates have 2 and 2 pairs, respectively. The 2 isolates have same number, 16 and 8, of submetacentric and subtelomeric pairs [13].Genetic relationships between geographical isolates have been studied by employing sequence analyses of nuclear ribosomal DNA (18S, internal transcribed spacer 1 and 2; ITS1 and ITS2) and mitochondrial DNA (cytochrome c oxidase subunit 1 [CO-1]). C. sinensis Korean isolates collected in Kimhae city were compared with Chinese isolates collected in Shenyang, Liaoning and Nanning, and Guangxi provinces. The geographical isolates were nearly identical in terms of nuclear ribosomal and mitochondrial DNA sequences, and revealed no more than a 1% sequence difference between Korean and Chinese isolates [14][15][16][17]. Isozyme electrophoresis had been used to study trematode systematics, and isozymes of the Korean and Chinese isolates show homozygous monomorphic banding patterns. Furthermore, alpha-glycerophosphate dehydrogenase produced a unique band pattern and was found to differentiate geographical isolates of C. sinensis [14]. However, in C. sinensis, intraspecific genetic variations among geographical isolates in the SinoKorea region appear minimal.As compared with O. viverrini, C. sinensis ITS2 revealed 95% identity and differences at 28 nucleotide points. Furthermore, the mitochondrial CO1 gene of O. viverrrini was found to be 96% identical with that of C. sinensis. Even a different genus in the same family Opisthorchidae, namely, O viverrrini appears to be genetically close to C. sinensis [17].

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“…Clone CsHA-16 (Cs28GST) cDNA was 799 bp long and encoded a polypeptide of 212 amino acids showing sequence identity (25-43%) with the 28 kDa glutathione S-transferase of vertebrates and invertebrates, and which was identical to that of C. sinensis . Clone CsHA-19 (CsRP) cDNA was 844 bp long and encoded a full open reading frame that shared homology with the repetitive proteins of invertebrates, including C. sinensis (Kim et al 2001). Clone CsHA-24 (CsMCP) harbored a 1,525-bp long cDNA encoding a polypeptide that showed a low degree of homology (~27%) with migratory cell-specific protein.…”

Section: Antigenic Clonesmentioning

confidence: 99%

Multiple recombinant antigens of Clonorchis sinensis for serodiagnosis of human clonorchiasis

Shin

Cho

et al. 2010

Parasitol Res

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Antigenic proteins from Clonorchis sinensis have been previously purified and evaluated for their antigenicity to enable the serodiagnosis of clonorchiasis. Though they were of high specificity, molecularly defined proteins were reported to be less sensitive as single antigens than crude antigen. To resolve this issue, 11 clones were selected by immunoscreening an adult C. sinensis cDNA library using infected human sera. Mixed antigens were prepared using recombinant proteins of positive clones and investigated for antigenicity by immunoblotting against C. sinensis- and helminth-infected patient sera. A mixed antigen of recombinant 28 and 26 kDa glutathion S-transferases (Cs28GST and Cs26GST) produced 76% sensitivity and 95% specificity. Furthermore, a triple mix of recombinant Cs26GST and Cs28GST with vitelline precursor protein pushed up the sensitivity to 87% and maintained specificity at 95%. It is proposed that multiple antigen mixes should be further studied to develop rapid serodiagnostic test kits for the serodiagnosis of human clonorchiasis.

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Molecular cloning and characterization of an antigenic protein with a repeating region from Clonorchis sinensis

Cited by 18 publications

References 32 publications

Analysis of the genes expressed in Clonorchis sinensis adults using the expressed sequence tag approach

Analysis of the genes expressed in Clonorchis sinensis adults using the expressed sequence tag approach

Functional Genes and Proteins of Clonorchis sinensis

Multiple recombinant antigens of Clonorchis sinensis for serodiagnosis of human clonorchiasis

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