人β_2m转基因小鼠的制备及鉴定(英文)

 制备人 β2m转基因小鼠 ,研究HLA B2 70 4基因的表达 .应用显微注射将人 β2m基因注入C5 7BL 6×昆明鼠和昆明鼠×昆明鼠F1代受精卵 .出生动物及其后代经PCR筛选 ,采用斑点杂交和Southern杂交对阳性鼠基因组DNA标本进行进一步鉴定和测定整合拷贝数 ,利用RT PCR检测阳性鼠中人 β2m转基因的表达 .6只原代仔鼠及 7只它们的下一代鼠 (F1)带有人 β2m基因 .由微注射基因后移卵出生的 86只小鼠中 ,C5 7BL 6×昆明鼠杂交仔鼠 35只 ,其中 4只阳性 (11 4 % ) ,昆明鼠×昆明鼠杂交仔鼠 5 1只 ,其中 2只阳性 (3 9% ) ,含有人 β2m基因的原代鼠×昆明鼠杂交仔鼠 2 0只 ,其中 7只阳性 .整合的转基因均为单拷贝 .Southern杂交证实上述阳性鼠确有转基因整合 .阳性鼠的皮肤、结肠、睾丸和脾脏组织中均有人β2m转基因mRNA的表达 .在转基因动物制备中 ,C5 7BL 6×昆明鼠F1代明显优于昆明鼠×昆明鼠F1代 .与人HLA B2 70 4基因相比 ,人 β2m基因不易整合 ,其整合率与整合拷贝数均较低 .得到的人 β2m转基因小鼠能够将人 β2m基困传给下一代 ,并可与人HLA B2 70 4转基因鼠交配 ,研究它的致病性

Production and Characterization of Human Microglobulin Transgenic Mice

The human β2 microglobulin transgenic mice were produced for the study of human HLA B 2704 gene related to arthrospondylosis in chinese.Human β2 microglobulin gene (β2m gene) was introduced into fertilized eggs of C57BL/6×Kunming and Kunming×Kunming F1 mice by microinjection.By PCR,the founder mice and their progeny were screened for integration of the human β2m transgene into the mouse genome which were further confirmed by Southern blotting.The copy number of transgene was determined by dot blotting.The expression of human β2m transgene in mice at the mRNA level was analyzed by RT\|PCR.Six founder mice and seven founder F1 contained human β2m gene.Four of 35 (11\^4%)hybrid mice from C57BL/6×Kunming and two of 51 (3\^9%) hybrid mice from Kunming×Kunming background carried human β2m gene.Statistical analysis showed that there was significant difference of the integration rate between the two groups( P <0 05),seven of 20 hybrid mice from founder mice containing human β2m gene×Kunming background carried human β2m gene.The transgene was the single copy in all the founder mice and founder F1.The human β2m gene was expressed at the mRNA level in skin,colon,testicle and spleen tissues of the transgenic mice.Human β2m transgenic mice have been obtained.For the production of transgenic mice,C57BL/6×Kunmin F1 is better than Kunming×Kunming F1.Compared with the HLA B 2704 gene,the integration of human β2m gene into mouse genome is more difficult.