| 1. | Then we transfected the plasmid to the bmn cells through lipofectin
在不同的时间，收集细胞上清。 |
| 2. | ( 2 ) the chromosomes of bmn cells showed the typical characteristics of lepidoterati insects . chromosomes of matephase without apparent centromere are short pole - like and pellet - like
( 2 ) bmn细胞的染色体表现为典型的鳞翅目昆虫的染色体：中期染色体短杆状或颗粒状、无明显的着丝粒。 |
| 3. | The bmn cells are multiploidy and heteroploidy so highly that in which the diploid was few and far between , and multiploid was so much that came to 90 % . even there were 533 chromosomes in one cell
Bmn细胞是高度异倍化、高度多倍化的细胞系，其典型二倍体极少而高多倍化群达90 ，最多的染色体一群在500条以上。 |
| 4. | ( 4 ) although the majority of bmn cells " mitoses are normal mitoses , there are still some abnormal phenomena found such as analogous amitoses , mitoses in multiploid cells and endomitoses
K ） bmn细胞有丝分裂以正常有丝分裂为主，但同时存在多倍核有丝分裂、核内有丝分裂以及类似于缢缩和劈裂等的类无丝分裂等现象。 |
| 5. | We generated a recombinant eukaryotic gene expressing vector harboring a full - length hgh gene , 2 . 4 kb genomic dna with four introns and the signal peptide sequence cloned to the eukaryotic gene expressing vector pcdna3 . 0
我们直接将含有4个内含子和自身信号肽的2 . 4kb全长人生长激素基因直接克隆至真核表达载体pcdna3 . 0 ，然后利用脂质体转染家蚕bmn细胞，瞬时表达hgh 。 |
| 6. | The recombinant transfer vector pbacpak - hbmp was constructed by insertion of the hbmp coding sequences into the multiple cloning site of transfer vector pbacpak . 8 . bmn cell line was co - transfected with pbacpak - hbmp plasmid and linearized baculovirus bacpak6 dna by dosper agent
将克隆到的hbmp基因通过适当的酶切插入到转移载体质粒pbac - pak8的多克隆位点中，获得重组转移载体质粒pbacpak - hbmp 。 |
| 7. | Expression amount of rhbsag in the former was increased 60 % and 80 % respectively than the latter . antigenicity of pres2 of fusion protein was increased 8 times and 10 times in bmn cells and in pupa hemolymph respectively than nonfusion hbsag
Bmpak - hbmp表达的融合m蛋白( pres2 + s )中， pres2抗原性显著提高，与bmpak - hbm表达的非融合蛋白相比，融合蛋白pres2抗原性在细胞中约提高了8倍，蛹中约提高了1o倍。 |
| 8. | The reasults are summed up as following : ( 1 ) the more metaphase chromosomes of bmn cells and moderately contraction can be obtained by air - drying after cultured at the colchicine concentration 1 . 0 u g / ml , 28 " c , for 6 - 9h
用空气干燥法和培养细胞贴片法对bmn细胞的染色体和有丝分裂进行研究发现： ( 1 )采用空气干燥法制备bmn细胞的染色体，在终浓度0 . 1ug / ml的秋水仙素28培养6 9h条件下可得到较多中期分裂相，染色体的凝集也比较适中。 |
| 9. | During the different time point , we retrieve the cell supernatant and through the electrophoretic analysis and immunoblot of the recombinant protein indicated that the cell which transfected the recombinant eukaryotic gene expressing vector could synthesis and excrete the peptide effectively
提取培养细胞总rna反转录cdna测序结果表明人生长激素基因组dna在家蚕bmn细胞内能正确转录，剪接。蛋白质电泳分析和免疫学检测证明转染细胞能够有效合成并分泌hgh蛋白质。 |
| 10. | The reasults are summed up as following : 1 the study on chromosomes and mitoses of bmn cells the cell line , bmn , is a silkworm cell line widely used in silkworm molecular genetics , cell engineering , gene engineering and baculovirus expression system but whose genetics and cytobiology studies are nearly untouched . the chromosomes and mitoses of the bmn cells are researched by the air - drying method and culturing cells on cover glasses
同时，还通过原代培养实验对新的家蚕胚胎细胞系的建立进行了探索和尝试，并对家蚕胚胎原代培养过程中出现的细胞和组织类型进行了观察、探讨与研究。 1bmn细胞有丝分裂及染色体研究bmn细胞是家蚕分子遗传学，细胞工程、基因工程和杆状病毒表达系统中广泛应用的家蚕细胞，但其遗传学和细胞生物学背景知之甚少。 |
