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Isolation of genes that are induced by Ty21a in intestinal cells by suppression subtractive hybridization (SSH)

Jie Gao

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Abstract

Objective:To isolate the genes induced by Ty21a. Methods: Prepare the intestinal cells of Balb/C mouse with and without Ty21a immunization, extract the polyA+ RNA, synthesize the dscDNA by using reverse transcription-PCR,get the differentially expressed genes by suppression subtractive hybridization(SSH).Results:Seven genes were novel genes revealed by GenBank searching whose expression were probably induced by Ty21a were isolated by SSH and cDNA microarray. The full length cDNA sequence of three genes are now obtaining through RACE-PCR. Conclusion: SSH is an effective method for isolating the differentiatlly expressed genes; High throughput cDNA microanay is a credibility method to profile changes in gene expression; Ty21a can induce the expression of some new genes related to immune.

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Objective:To isolate the genes induced by Ty21a. Methods: Prepare the intestinal cells of Balb/C mouse with and without Ty21a immunization, extract the polyA+ RNA, synthesize the dscDNA by using reverse transcription-PCR,get the differentially expressed genes by suppression subtractive hybridization(SSH).Results:Seven genes were novel genes revealed by GenBank searching whose expression were probably induced by Ty21a were isolated by SSH and cDNA microarray. The full length cDNA sequence of three genes are now obtaining through RACE-PCR. Conclusion: SSH is an effective method for isolating the differentiatlly expressed genes; High throughput cDNA microanay is a credibility method to profile changes in gene expression; Ty21a can induce the expression of some new genes related to immune.

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Available abstract

Objective:To isolate the genes induced by Ty21a. Methods: Prepare the intestinal cells of Balb/C mouse with and without Ty21a immunization, extract the polyA+ RNA, synthesize the dscDNA by using reverse transcription-PCR,get the differentially expressed genes by suppression subtractive hybridization(SSH).Results:Seven genes were novel genes revealed by GenBank searching whose expression were probably induced by Ty21a were isolated by SSH and cDNA microarray. The full length cDNA sequence of three genes are now obtaining through RACE-PCR. Conclusion: SSH is an effective method for isolating the differentiatlly expressed genes; High throughput cDNA microanay is a credibility method to profile changes in gene expression; Ty21a can induce the expression of some new genes related to immune.

Key concepts: Suppression subtractive hybridization, Gene, Biology, Complementary DNA, Gene expression, GenBank, Genetics, Molecular biology

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Isolation of genes that are induced by Ty21a in intestinal cells by suppression subtractive hybridization (SSH) — Research Paper | ScholarLens