文档介绍：摘 要
本研究旨在探索HBV S/HEV ORF2融合基因在植物中表达乙肝戊肝双价抗原的可能性,以期为进一步研制植物乙肝戊肝双价口服疫苗打下基础。
应用重组PCR技术将乙肝病毒(HBV)S基因连接在戊肝病毒(HEV)ORF2部分序列的5’端,二者通过柔性肽(Gly。Ser):序列相连,构
建成融合基因BE,测序结果正确。
将融合基因BE克隆到植物双元表达载体pBin438上,获得 pBin438BE,然后用冻融法将其转入根癌农杆菌EHAl05中,侵染丽春番茄子叶和下胚轴,经预培养、共培养、筛选培养、生根培养获得了5株抗卡那霉素的番茄植株。提取抗性植株基因组DNA,进行PCR及 PCR—Southern检测,4株获得阳性信号。初步表明目的基因已整合到了番茄基因组中。
关键词:乙肝病毒s基因 戊肝病毒ORF2
esculentum Mill.)融合基因 农杆菌 转基因植物
ABSTRACT
T11iS research was to explore the feasibility of expressing HBV S脏V ORF2 fusion gene in plants for the purpose of developing a new type of hepatitis bivalent edible ine.
Hepatitis BⅥrus S gene Was connected to the 5’end of Hepatitis E Virus
ORF2 partial sequence with the technique of binant designed DNA sequence which encoded a polypeptide linker(Gly4Ser)2 Was used to
splice the two sequencing results showed that the fusion
gene(BE)was essfully constructed.
rnle fusion gene(BE),approximatelyl 300bp,Was cloned to the plant binary
expression vector plasmid pBin438BE under the control of DE35S promoter Was constructed and transformed into Agrobactrium mmefaciens E}王A1 05 via freeze-thaw explants from cotyrledons and hypocotyls were cultured essively on
medium、 CO—cultivation medium、 selective medium and
root-growth media 5 Km-resistant tomato plants were regenerated plantlets were analyzed further by PCR and results confirmed that the fusion gene had been integrated into
tomato genome.
Key words:Hepatitis B Virus S gene Hepatitis E Virus ORF2
Tomato(Lylopersicon esculentum Mill.) Fusion gene
、Agrobactrium tumefaciens Transgenic plants
长春理工大学硕士学位诠支原刨性声明
本人都重声明:所呈交的硕士学位论又,{HBV S/IqEV ORF2融合基因的构建及对番茄的转化》是本人在指导教师的指导下,独立进行研究工作所取得的或果。豫文孛已经注疆弓l用韵内容秀,本论文不包含任俺其纯令人或集体已经发袭或撰写过的作品成果。对本文的研究做出重要贡献的个人和集体,均己在
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作者签名:至蒌巫五年』z胄2篁舀
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