Objective:To modify human α-lactalbumin yeast artificial chromosome(HLA-YAC)and to use it as vectors for analysis of gene expression and regulatory elements of α-lactalbumin in vivo in the future.
目的?对人乳白蛋白酵母人工染色体(HLA-YAC)进行定点改造,以便进一步利用该载体作与基因表达调控相关的 研究。
Methods Three clones of yeast artificial chromosomes (YAC) related to bcr gene were examined by FISH in combination with Alu-PCR to identify metaphases peripheral lymphocytes from normal inpiduals and metaphases bone marrow cells from CML patients with chromosome t (9; 22).
方法应用Ａｌｕ－ＰＣＲ和荧光原位杂交（ＦＩＳＨ） 技术，对３个酵母人工染色体（ＹＡＣ）候选克隆进行鉴定，制备ＤＮＡ探针，并经正常人外周血淋巴细胞和慢性髓细胞性 白血病患者骨髓细胞筛选。
Yeast Artificial Chromosome(YAC)vector was modified with a CaMV35S promoter-GUS-NOSterminator or hygromycin genes,which is able to express in plants.
将能在植物中表达的烟草花叶病毒ＣａＭＶ３５Ｓ启动子、ＧＵＳ基因、潮霉素（Ｈｙｇｒｏｍｙｃｉｎ）基因及ＮＯＳ终止子组建到酵母人工染色体（ＹＡＣ）载体，构建了具有能在植物细胞中表达和选择ｐＹＡＶ、ＧＵＳ、ｐＪＳ９７／ｐＪＳ９８－ＨＹＹＡＣ载体系统和一个标记ＹＡＣ克隆的ｐＵＲ４３－ＨＹ质粒。
There were some disadvantages on yeast artificial chromosome (YAC) including low cloning efficiency, high incidence of chimeric clones, instability and deletions of insert DNA, but YAC took large DNA inserts that was not able to replace by the other vector, which made YAC to be one of the important tool in study of human and animal genome.
尽管酵母人工染色体 (Yeast artificial chrom osome,YAC)存在着克隆效率低、嵌合体出现率高、部分克隆不稳定和缺失等弊端 ,但由于它容纳高分子量 DNA的优点其它载体无法替代 ,而成为人类和动物基因组研究中最为主要的研究工具之一。