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[Enrichment and screening of up-regulated genes of the mosquito Anopheles stephensi in response to malaria parasite].

Abstract

OBJECTIVE

To isolate and identify genes related to malaria parasite infection in vector mosquito, and to explore the mechanisms.

METHODS

Anopheles stephensi infected with Plasmodium yoelii was used as tester (T) group, while uninfected but normal blood fed as driver (D) one. Engorged female mosquitoes of two groups were collected separately at 24 hours after biting. An enriched subtractive cDNA pool was generated through the course of suppression subtractive hybridization (SSH) and selective PCR amplification. The subtracted library was screened by hybridization using T and D cDNA mixture as probes, respectively. The positive clones, which produced stronger signal when probed with T than with D, were sequenced and their sequence homologues in GenBank database were searched with BLAST by internet.

RESULTS

The analysis of subtraction efficiency showed that the differentially expressed genes in T comparing to in D were enriched significantly. In dot blot screening, 24 of 58 randomly selected clones (41.4%) were shown up-regulation in malaria infected mosquitoes. The BLAST search of 23 genes revealed that 12 were homologous to functionally known genes, 4 were homologous to functionally unknown entries, and 7 were novel without any relatives. Nine of the 23 genes (39.1%) also hit homologous sequences in the An. gambiae EST database generated from an immune competent cell line treated with lipopolysaccharide (LPS).

CONCLUSION

An enriched cDNA pool of the mosquito genes which up-regulated responsively at the early stage of malaria parasite infection was obtained. Expression screening against the pool indicated that various biochemical processes and mechanisms might be involved in the response of mosquito to parasite infection, especially those related with the innate immune system and energy metabolism.

Authors+Show Affiliations

Department of Etiologic Biology, Second Military Medical University, Shanghai 200433.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

English Abstract
Journal Article
Research Support, Non-U.S. Gov't

Language

chi

PubMed ID

12572060

Citation

Xu, X C., et al. "[Enrichment and Screening of Up-regulated Genes of the Mosquito Anopheles Stephensi in Response to Malaria Parasite]." Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi = Chinese Journal of Parasitology & Parasitic Diseases, vol. 19, no. 6, 2001, pp. 325-9.
Xu XC, Qu FY, Song GH, et al. [Enrichment and screening of up-regulated genes of the mosquito Anopheles stephensi in response to malaria parasite]. Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2001;19(6):325-9.
Xu, X. C., Qu, F. Y., Song, G. H., & Xu, J. N. (2001). [Enrichment and screening of up-regulated genes of the mosquito Anopheles stephensi in response to malaria parasite]. Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi = Chinese Journal of Parasitology & Parasitic Diseases, 19(6), 325-9.
Xu XC, et al. [Enrichment and Screening of Up-regulated Genes of the Mosquito Anopheles Stephensi in Response to Malaria Parasite]. Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2001;19(6):325-9. PubMed PMID: 12572060.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - [Enrichment and screening of up-regulated genes of the mosquito Anopheles stephensi in response to malaria parasite]. AU - Xu,X C, AU - Qu,F Y, AU - Song,G H, AU - Xu,J N, PY - 2003/2/8/pubmed PY - 2003/10/28/medline PY - 2003/2/8/entrez SP - 325 EP - 9 JF - Zhongguo ji sheng chong xue yu ji sheng chong bing za zhi = Chinese journal of parasitology & parasitic diseases JO - Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi VL - 19 IS - 6 N2 - OBJECTIVE: To isolate and identify genes related to malaria parasite infection in vector mosquito, and to explore the mechanisms. METHODS: Anopheles stephensi infected with Plasmodium yoelii was used as tester (T) group, while uninfected but normal blood fed as driver (D) one. Engorged female mosquitoes of two groups were collected separately at 24 hours after biting. An enriched subtractive cDNA pool was generated through the course of suppression subtractive hybridization (SSH) and selective PCR amplification. The subtracted library was screened by hybridization using T and D cDNA mixture as probes, respectively. The positive clones, which produced stronger signal when probed with T than with D, were sequenced and their sequence homologues in GenBank database were searched with BLAST by internet. RESULTS: The analysis of subtraction efficiency showed that the differentially expressed genes in T comparing to in D were enriched significantly. In dot blot screening, 24 of 58 randomly selected clones (41.4%) were shown up-regulation in malaria infected mosquitoes. The BLAST search of 23 genes revealed that 12 were homologous to functionally known genes, 4 were homologous to functionally unknown entries, and 7 were novel without any relatives. Nine of the 23 genes (39.1%) also hit homologous sequences in the An. gambiae EST database generated from an immune competent cell line treated with lipopolysaccharide (LPS). CONCLUSION: An enriched cDNA pool of the mosquito genes which up-regulated responsively at the early stage of malaria parasite infection was obtained. Expression screening against the pool indicated that various biochemical processes and mechanisms might be involved in the response of mosquito to parasite infection, especially those related with the innate immune system and energy metabolism. SN - 1000-7423 UR - https://www.unboundmedicine.com/medline/citation/12572060/[Enrichment_and_screening_of_up_regulated_genes_of_the_mosquito_Anopheles_stephensi_in_response_to_malaria_parasite]_ DB - PRIME DP - Unbound Medicine ER -