Generation of Birds Maintaining Identical Genome

维持相同基因组的鸟类的产生

基本信息

批准号：
15380190
负责人：
KAGAMI Hiroshi
金额：
$ 10.3万
依托单位：
Shinshu University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2005
项目状态：
已结题

项目摘要

Fertilized eggs of the chicken were obtained. The egg shells were cracked on Petri dish. Most of the albumen around the egg yolk was removed. A filter paper was put around an embryo. The egg membrane around the filter paper was cut by a surgical scissors. The blastoderm alone was obtained. The developmental region either area pellucida or area opaca was identified under a microscope. Pluripotent cells were isolated from a blastoderm of the chicken. The cells from a blastoderm (donor) were injected into recipients' male or female blasoderms to generate germline chimeras. The donor strains were either Barred Plyomouth Rock (BPR) or Silky. The recipient strain was White Leghorn. The donor cells were injected into either male or female recipient embryos. The manipulated chimeric embryos were cultured by ex vivo culture system. The somatic chimerism were accessed according to the black pigmentation due to the donor feather. The hatched chimeras were raised until sexual maturities. Three pairs of the male and female somatic chimeras derived from a blastoderm were generated according to the present experimental strategies. A pair of male and female chimeras reached sexual maturity. When the male and female were test mated, one offspring which had black spot derived from donor strain (BPR) in the feather was generated. It was suggested that sperm or ova was generated from the donor cells. The developed strategy should be powerful tool for generation of birds maintaining identical genome (similar to clone birds) in the future.

获得了鸡肉的受精卵。在培养皿上将蛋壳破裂。蛋黄周围的大多数蛋白被去除。将滤纸放在胚胎周围。滤纸周围的鸡蛋膜被外科剪刀切割。仅获得了胚泡。在显微镜下鉴定出卵石或Opaca区域的发育区域。从鸡肉的胚胚中分离多能细胞。将来自胚胚（供体）的细胞注入接受者的雄性或雌性蓝皮士，以产生种系嵌合体。供体应变要么被禁止普利茅斯岩石（BPR）或丝滑。接收者应变是白色的leghorn。将供体细胞注入男性或女性受体胚胎中。通过离体培养系统培养操纵的嵌合胚。根据供体羽毛而导致的黑色色素沉着，可以进入体细胞嵌合。将孵化的嵌合体抬高直至性成熟。根据目前的实验策略，产生了源自胚泡的三对雄性和女性体细胞嵌合体。一对雄性和雌性嵌合体达到了性成熟。当男性和雌性进行测试交配时，产生了羽毛中的供体菌株（BPR）的黑点的后代。有人提出，精子或OVA是由供体细胞产生的。开发的策略应该是生成鸟类的强大工具，以维持相同的基因组（类似于克隆鸟类）。