Lurquin P F, Behki R M
Mutat Res. 1975 Jul;29(1):35-51. doi: 10.1016/0027-5107(75)90019-6.
Escherichia coli [3H]DNA supplied to vegetative cultures of wild-type (mt+) and CW15 (mt+;mutant lacking the cell wall) Chlamydomonas reinhardi could bind to the cell wall of the wild-type and to the cell membrane of CW15 mutant cells. The extent of this binding decreased with time and was to a large degree (over 90%) DNA-ase-sensitive. Nevertheless, about 0.01% of the bacterial DNA remained irreversibly associated with the cells when they reached stationary phase. The irreversible binding of the donor bacterial DNA to Chlamydomonas cells could be increased by treatment of the cultures with polycations such as DEAE-dextran, poly-L-lysine and poly-L-ornithine. Although the CW15 cells rapidly degraded bacterial DNA in the culture medium wild-type cells showed only a small effect on the molecular weight of the donor DNA. The acid-insoluble radioactivity irreversibly bound to WT (+) cells consisted mainly of oligonucleotides with a small proportion present as less depolymerized donor DNA. No radioactivity, however, was found to be associated with the recipient high molecular weight Chlamydomonas DNA. No labeled donor DNA could be recognized in the cells given bacterial [3H]DNA in early stationary phase. Instead, radioactivity found in Chlamydomonas DNA corresponded to reutilization of [3H]thymine derivatives released as a result of [3H]DNA degradation. No evidence for the integration of detectable amounts of donor DNA sequences into the host cell DNA was obtained.
供给野生型(mt +)和CW15(mt +;缺乏细胞壁的突变体)莱茵衣藻营养培养物的大肠杆菌[3H] DNA能够与野生型的细胞壁以及CW15突变体细胞的细胞膜结合。这种结合的程度随时间下降,并且在很大程度上(超过90%)对DNA酶敏感。然而,当细胞进入稳定期时,约0.01%的细菌DNA仍不可逆地与细胞结合。用聚阳离子如DEAE - 葡聚糖、聚-L-赖氨酸和聚-L-鸟氨酸处理培养物,可以增加供体细菌DNA与衣藻细胞的不可逆结合。虽然CW15细胞能迅速降解培养基中的细菌DNA,但野生型细胞对供体DNA的分子量只有很小的影响。不可逆地结合到野生型(+)细胞上的酸不溶性放射性主要由寡核苷酸组成,只有一小部分以解聚程度较低的供体DNA形式存在。然而,未发现有放射性与受体的高分子量衣藻DNA相关。在稳定期早期给予细菌[3H] DNA的细胞中未检测到标记的供体DNA。相反,在衣藻DNA中发现的放射性对应于因[3H] DNA降解而释放的[3H]胸腺嘧啶衍生物的再利用。未获得可检测量的供体DNA序列整合到宿主细胞DNA中的证据。