摘要
目的探讨构建hSAMP32DNA疫苗的方法。方法根据GenbankhSAMP32cDNA序列设计引物,RT-PCR得到hSAMP32DNA,将其亚克隆至pGEM-T载体,酶切鉴定后测序;再构建pcDNA3.1(+)-hSAMP32质粒,并对质粒DNA定量。结果(1)人睾丸组织中提取的总RNA显示2条清晰条带,分别对应28S、18S;RT-PCR扩增产物与预期的目的基因hSAMP32长度一致;(2)RT-PCR产物与载体pGEM-T连接后测序,显示其与Genbank的hSAMP32基因序列一致;(3)超螺旋的pcDNA3.1(+)-hSAMP32DNA疫苗不含内毒素。结论应用亚克隆技术构建hSAMP32DNA避孕疫苗的载体pcDNA3.1(+)-hSAMP32,为研制避孕疫苗奠定实验基础。
Objective To establish the DNA contraceptive vaccine of hSAMP32. Methods The hSAMP32 DNA was acquired by RT-PCR based on the hSAMP32 sequence in Genbank, and subcloned into pGEM-T-vector; and the sequence was detected after digestion. Then the pcDNA3.1 （＋）-hSAMP32 plasmid DNA was constructed and quantified. Results （1）The distinct total RNA bands corresponding to 28 S and 18 S clearly were isolated from human testis tissue. The length of amplified RT-PCR product was consistant with that of proposed target gene, hSAMP32. （2）The sequence of RT-PCR product linked with pGEM-T vector was co-incident with that of hSAMP32 in Genbank. （3） The suporeoiled pcDNA3.1 （＋）-hSAMP32 contained no endotoxin. Conclusion Establishment of DNA immuno-vaccine pcDNA3.1 （＋）-hSAMP32 by using subcloning technique may provide experimental baed for research of contraceptive vaccine.
出处
《解剖学研究》
CAS
2006年第3期209-212,共4页
Anatomy Research
关键词
精子
顶体
免疫避孕
DNA避孕疫苗
Sperm
Acrosome
Immuno-contraception
DNA contraceptive vaccine
