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Study on the Expression of Partial Different Genes in Segments about Moniezia

Author: WangLiXiao
Tutor: BaoXinWen
School: Shihezi University
Course: Preventive Veterinary Medicine
Keywords: Moniezia mRNA sequence analysis mRNA Differential expression analysis Prokaryotic expression of membranin-like protein
CLC: Q78
Type: Master's thesis
Year: 2010
Downloads: 5
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Abstract


Five M. expansa’s genes were studyed that the expression abundance was large and had a significant differences in each stage which got through the gene plasmid library. Gene plasmid library numbers were SSBR88,SSBR116,SSBR126,SSBR210,SSBR614.We got five full length mRNA sequences after the genes were cloned and sequenced.After that we sent them on the GenBank,and analysed them with blast on line.GenBank accession number were GO254238-GO254242.Later genes were analyzed with bioinformatics.Simultaneously the genes’differential transcription was investigated in each developmental stage of two kinds of Moniezia’s scolex,immature proglottid,mature proglottid and gravid proglottid with SYBR Green real-time PCR detection method.Finally,the specific gene of M. expansa which had a high expression in gravid segment which similar to the membrane protein was expressed and analyzed by constructing the expression vector.The sequence analysis results showed that we had successfully obtained the full-length mRNA sequences of the five genes.SSBR116,SSBR210 were the highest similarity of 74.8% and 80% respectively with the Fasciola’s enolase and Schistosoma japonicum’s P-tubulin through the BlastX.The two genes were initially identified for the enolase and P-tubulin gene.of M.expansa.The other three genes preliminary judged as the specific genes of M. expansa for they had a low similarity with the genes in the database.According to the protein structure,they were respectively inferenced as the similar genes with membrane,antigen and chorionic.Real time RT-PCR results showed that the tendency of the two cestode’s mRNA expression discrepancy were at a equal pace.The mRNA expression abundance of enolase (SSBR116) in each segment was notable different,they were mature proglottid,immature proglottid,gravid proglottid and scolex from high to low on expression abundance.lt suggested that the enolase gene as the main enzymes of glycometabolic,it played a key role for the process of cell metabolism and energy generation in their life.The mRNA expression abundance ofβ-tubulin in the mature proglottid was highest.As the basic unit of the spindle structur during the process of cell division and the cytoskeletal protein.It suggested us that the gene might be playing a key role to maintain the structure of cellular tissue,cellular function,cell division and gametogenesis in their life.The unknown protein SSBR88 was similar to membrane protein,its mRNA expression abundance was highest in the gravid proglottid. Suggested it might be correlating with the formation of egg membrane in the ovification.In addition,SSBR126 as a new antigen-like protein,Its mRNA expression abundance was highest in the gravid proglottid too.and its role needs further experimental verification.SSBR614 had some similarity with chorionic protein of mice.Its mRNA expression abundance was highest in the mature proglottid.Suggested it was concerned with the formation of the.female reproductive organ.In the paper,to exploring mechanism of molecular action about growth and development of Moniezia,five differential expression genes was cloned and analyzed with bioinformatics through the the M.expansa’s gene plasmid library.Simultaneously the genes’differential expression was investigated in each developmental stage of two kinds of Moniezia with SYBR Green real-time PCR detection method.Besides,the study on the expression about the analogous gene of membrane protein offered a reference for lucubrating the gene’s biological function from M. expansa.

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CLC: > Biological Sciences > Molecular Biology > Genetic engineering (genetic engineering)
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