西安交通大学学报（医学版）2011，Vol.32Issue(1)：131-134,4.
Tex13基因非放射性原位杂交探针的制备
Preparation of non-radiolabeled in situ hybridization probes of Tex13 gene
摘要
Abstract
Objective To prepare digoxigenin ( DIG )-labeled non-radioactive in situ hybridization cRNA probes of testis expressed gene 13 (Tex13 gene) in order to determine Tex13 gene expression in the mouse gonad.Methods RT-PCR was performed for amplification of Tex13 gene, and the PCR fragments were cloned into the polylinker site of the pGEM-T Easy vector which contained a promoter for SP6 and T7 RNA polymerase. DH10B competent cells were used for transformations and the recombinant clones were identified by blue/white screening.The recombinant plasmids served as templates for generation of PCR products of Tex13 gene with the SP6 and T7 primer. We used recovered PCR products to synthesize DIG-labeled anti-sense and sense RNA probes by in vitro transcription with SP6 and T7 RNA polymerases and DIG RNA Labeling Mix. Results Non-radioactive in situ hybridization studies on the expression of Tex13 gene in the normal mouse testis showed that the probes had relatively high specificity and sensitivity. Conclusion The successful preparation of DIG-labeled probes of Tex13 gene lays an experimental foundation for detecting the developmental expression pattern of Tex13 gene in the mouse gonad and studying its role in the meiosis.关键词
Tex13/生殖细胞/减数分裂/RNA探针/原位杂交分类
生物科学引用本文复制引用
霍涌玮,邱曙东,周庆,李莹,聂蓉,FRIEL P,MITCHELL D,SMALL C,GRISWOLD MD..Tex13基因非放射性原位杂交探针的制备[J].西安交通大学学报（医学版）,2011,32(1):131-134,4.基金项目
陕西省自然科学基础研究计划项目(No.SJ08C210) （No.SJ08C210）
