本文選題：牛支原體 + 克隆��； 參考：《內蒙古農業(yè)大學》2015年碩士論文

【摘要】：牛支原體是能夠導致牛肺炎、關節(jié)炎、流產、乳腺炎等多種疾病的一種病原體。給養(yǎng)牛業(yè)造成了巨大的經濟損失,嚴重影響了養(yǎng)牛業(yè)的健康發(fā)展。由于王艷杰測序的M. bovis NM 2012菌株的全基因組序列存在序列缺口和多余的重復序列,為了得到M.bovis NM 2012基因組完整序列,本試驗對王艷杰測序的M.bovis NM 2012進行基因組補缺口(補洞)工作。本試驗選擇M.bovis HB0801為參考基因序列,經過將王艷杰測序的M.bovis NM 2012與M.bovis HB0801比對后發(fā)現,共有34處未完成的基因序列,在未完成的序列兩端各增加200～1,000bp設計引物,進行PCR擴增和克隆測序,并利用生物信息學軟件進行拼接,從而完成M.bovisNM 2012菌株的全基因組補缺口(補洞)工作,并應用生物信息學軟件對M.bovisNM 2012菌株的ORFs、脂蛋白、IS元件等進行預測,并對相關的基因進行功能注釋。比對結果顯示,在34處未完成的基因序列中,有21處由字母N代替,字母N的長度是在10～2,359bp之間,字母N的總長度為13,131bp；34處未完成的序列中,有23處是缺口序列,總共缺失的序列是24,763bp,11處多余重復的序列長度為14,765bp�？寺y序結果表明,在23處缺口序列中,總共缺失的序列是20,222bp；11處多余重復序列的總長度為14,765bp,實際有10,226bp為無效的重復序列,4,539bp確為M.bovis NM 2012本身的序列；本試驗中,去除由未知序列N代替的13,131bp和無效的重復序列10,226bp,補齊缺失的序列20,222bp。將王艷杰測序的全基因組大小為993,483bp的M.bovis NM 2012,經過補缺口工作,最終拼接得到M. bovis NM 2012全基因組大小為990,348bp。M.bovis NM 2012菌株全基因組大小為990,348bp, GC%含量為29.29%;與M.bovis HB0801進行比對后,預測編碼區(qū)含839個ORFs,含有34個tRNA,6個rRNA,預測含有103個脂蛋白和2個Vsp。通過對5株牛支原體全基因組進行基因組共線性分析比較,M. bovis NM 2012基因組與M.bovis PG45基因組的相似性最小,與M.bovis HB0801基因組的相似性最高。
[Abstract]:Mycoplasma bovis is a pathogen that can cause many diseases, such as pneumonia, arthritis, abortion, mastitis and so on. It has caused enormous economic losses to the cattle industry and seriously affected the healthy development of the cattle industry. In order to obtain the complete genome sequence of M.bovis NM 2012, there is a gap in the whole genome sequence and redundant repeat sequence in the whole genome sequence of M. bovis NM 2012, which was sequenced by Wang Yanjie. M.bovis NM 2012, which was sequenced by Wang Yanjie, was studied in this experiment. In this experiment, M.bovis HB0801 was selected as the reference gene sequence. After comparing the M.bovis NM 2012 and M.bovis HB0801 sequenced by Wang Yanjie, it was found that there were 34 uncompleted gene sequences, and the primers designed by 200~1000bp were added at each end of the unfinished sequence. PCR amplification, cloning and sequencing were carried out, and bioinformatics software was used to assemble the whole genome gap (hole) of M.bovisNM 2012 strain. Bioinformatics software was used to predict ORFs, lipoprotein is elements of M.bovisNM 2012 strain, and to annotate the related genes. The results showed that of the 34 uncompleted sequences, 21 were replaced by the letter N, the length of the letter N was between 10~2359bp, and the total length of the letter N was 13131bpng34, of which 23 were gap sequences. The total missing sequence was 24763bpmpn11, and the length of the redundant repeats was 14765bp. The results of cloning and sequencing showed that the total missing sequence of 23 gap sequences was 20222bp-1, the total length of redundant repeats was 14765 BP, and the 4539bp repeats with 10226bp as invalid were indeed the sequences of M.bovis NM 2012. The 13131bp replaced by the unknown sequence N and the invalid repeat sequence 10226bpwere removed, and the missing sequence 20222bpwas added. The whole genome size of M.bovis NM2012, which was 993483bp, was sequenced by Wang Yanjie. After filling the gap, the whole genome size of M. bovis NM 2012 strain 990348bp.M.bovis NM 2012 was 990348bpand the content of GC% was 29.290.After comparing with M.bovis HB0801, the whole genome size of M. bovis NM 2012 was 990348bpand the content of GC% was 29.29. The predicted coding region contained 839 ORFs, 34 tRNAs, 6 rRNAs, 103 lipoproteins and 2 Vsps. The genomic collinear analysis of 5 strains of Mycoplasma bovis was conducted to compare the similarity between M. bovis NM 2012 genome and M.bovis PG45 genome, and the highest similarity with M.bovis HB0801 genome.
【學位授予單位】：內蒙古農業(yè)大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S852.62

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