Research On The Major Genetic Characteristics Of In Vitro Cultured Fibroblast Cells Of Three Local Chinese Chickens

1. Fibroblast cell lines of White earlobes chicken, Wenchang chicken and Chinese gamecock chicken were successfully established by primary culture and serial subcultivation. The sample content was 57, 53 and 43 respectively. The number of frozen tube was 322, 237 and 258 respectively. The cell number per frozen tube was respectively 1.52×107, 2.67×107 and 3.41×107 cell/ml.2. The diploid cell in White earlobes chicken, Wenchang chicken and Chinese gamecock chicken is 86%, 92%, and 89% respectively by analyzing the chromosomal metakinesis. The diploid chromosomal number of three chickens is 2n=78, consist of 10 pairs of macrochromosomes and 29 pairs of microchromosomes, and sex chromosome type is ZZ(♂)/ZW(♀).3. By preparing the low ionic strength of polyacrylamide gel electrophoresis, the results of isoenzyme analysis is as follow: there are 4 enzymograms of LDH in three kind of chickens, which are LDH1, LDH2, LDH3 and LDH4 from anode to cathode. There is LDH5 in Wenchang chicken and Chinese gamecock chicken, which is illegible. The relative mobility are similar between 5 enzymograms. There are 2 enzymograms of MDH in three chicken's cells, and the relative mobility of m-MDH is faster than that of s-MDH. 4. The transfection condition is better optimized than before in this research. The optimized condition is obtained by transfecting recombinant plasmid pEGFP-C1 of 2μg with liposome of 6ul and exposing the plasmid-liposome complex to the fibroblast cells of Wenchang chicken about 48 hours. Transient transfection results manifest: at 48hr, pEYFP-N1 have the highest transfection efficiency in three chicken's cells, which are 46.52%, 58.63% and 49.23% respectively. Then the efficiency of pEGFP-N1 is 58.36% in Wenchang chicken. After 72hr, although the transfection efficiency has some degressive tendency (pEGFP-C1 in White earlobes chicken is exception), pEGFP-C1 keep a good expression of 55.63% in Wenchang chicken. The cytoplasm and caryon besides some capsular vesic are filled with green fluorescent protein. There is a circularity outline surrouding the caryon which consist of various granulo-expression product of pDsRed1-N1 and pECFP-N1. Transfected cells have a normal growth and proliferation.