鹌鹑肝组织胱硫醚β-合酶基因RT-PCR的实验研究  

An Experimental Study on RT-PCR for Cystathionine Beta-synthase Gene in Partridge Liver Tissue

作　　者：李丽帆[1] 彭义[1] 吴芳[1] 

机构地区：[1]广西中医学院,广西南宁530001

出　　处：《广西中医学院学报》2007年第4期6-9,共4页Journal of Guangxi Traditional Chinese Medical University

基　　金：广西科技厅资助课题(桂科自0339058)

摘　　要：[目的]建立一种有效的鹌鹑肝组织胱硫醚β-合酶(CBS)RT-PCR,为实现CBS Real-time PCR提供条件。[方法]采取一步法,改进一步法,异硫氰酸胍法(重复抽提)和异硫氰酸胍法(一步法)四种方法提取总RNA,以CBS cDNA共有序列设计鹌鹑CBS引物,进行CBS RT-PCR引物筛选。[结果]除改进一步法外,其他三种方法提取的总RNA完整性好;除异硫氰酸胍法(一步法)外,其他三种方法提取的总RNA的A260/A280都在1.8以上纯度。异硫氰酸胍法(重复抽提)提取的总RNA极少残留DNA。[结论]四种方法都能较好地扩增CBS片段,一步法最省时。但异硫氰酸胍法(重复抽提)抽提得到的RNA质量较好.[Objective]To testify and establish a effective RT-PCR for cystathionine beta-synthase （CBS） in partridge liver tissue, [Methods]Four methods were used in total RNA extraction： One-step Method, Improved One-step Method,Guanidinium Isothiocyanate Method （ repetitive extraction） and Guanidinium Isothioeyanate Method （ one-step ）. Meanwhile, partridge CBS primers were designed according to CBS eDNA consensus sequence. Then CBS gene RT-PCR were carried on to evaluate and tilt these primers. [Results]Except Improved One-step Method, total RNA extracted through three other methods were in a good integrality. Except Guanidinium Isothiocyanate Method（one-step）, total RNA extracted through three other methods were in a good purity（ A260/A280 〉 1.8）. DNA residua were little in RNA extracted through Guanidinium Isothiocyanate Method（repetitive extraction）. [ Conclusions] The four applied method have a good performance in CBS segment amplification. One-step Method does best in saving time, so does Guanidinium Isothiocyanate Method（repetitive extraction）in obtaining the best-quality RNA.

关 键 词：胱硫醚Β-合酶 RT—PCR 总RNA 鹌鹑 动脉粥样硬化 

分 类 号：Q522.6[生物学—生物化学]