Generalized Bacteriophage Transduction in Serratia marcescens

Tatiana V. Shirshikova, Olga V. Morozova, Leisan Kh. Kamaletdinova, Margarita R. Sharipova,

& Lydia M. Bogomolnaya

Introduction

Generalized transduction is a way of transferring a gene from one bacterium to another

using bacteriophages. Instead of a viral DNA, the DNA of the infected bacterial cell is carried

and transferred by the phages to another. This process is used by molecular biologists to study

bacteria such as pathogens. Serratia marcescens are human pathogenic bacteria resistant to

several antibiotics. Their strains are highly diverse and different, so one strain cannot represent

another. The researchers aimed to determine the transduction potential of the bacteriophages,

ΦOT8 and ΦIF3, in the SM6 and SR41-8000 strains of S. marcescens.

Materials and Methods

The two S. marcescens strains, SM6 and SR41-8000, were cultured and subcultured into

LB broth, while the two bacteriophages, ΦOT8 and ΦIF3, were serially diluted. The resulting

strains and phages were mixed and incubated at 30 and 37°C.

TT392 TolC-6His, a kanamycin-resistant strain, was cultured and introduced with ΦOT8

to create plaques. The top agar was then gathered to prepare a phage lysate strain.

SM6 was grown in an LB broth and concentrated ten times. It was then mixed with the

prepared phage lysate, incubated overnight, and plated on plates with kanamycin. The grown

bacteria were then streaked.

 Western blotting was done to observe the transductants for the possible appearance of

TolC-6His strain.

Results

After the incubation at 30 °C, the activity of the bacteriophage ΦOT8 caused small

plaques to appear on the lawn of both SM6 and SR41-8000 strains (Fig. 1a). Meanwhile, no

plaques appeared on the same plates at 37°C. Furthermore, ΦIF3 showed less prominent results

at both temperatures, and were therefore not used at the proceeding experiments.

Red colored bacterial colonies appeared on the plates with kanamycin, while non in the

parent strain. Moreover, TolC-6His protein bands were observed in the SM6 strain through

Western blotting (Fig. 1b).

Fig1. ΦOT8 bacteriophage activity was present in both SR41-8000 and SM6 strains of S.

marcescens. a ΦOT8 caused formation of plaque (indicated by white arrows) at 30°C. b TolC-

6His protein bands were observed. Strains 1,2 ( SM6 TolC-6His), 3 (TT392,negative control),

and 4 (TT392 TolC- 6His) were used.

Discussion

The appearance of plaques in the bacterial lawn of SM6 and SR41-8000 strains showed

the range of the bacteriophage activity in each strain. Both ΦOT8 and ΦIF3 can cause cell lysis

of the two Serratia marcescens strains.

The appearance of the red-colored colonies and the TolC-6His protein band formation

indicated that transduction has occurred. The bacteriophages successfully transferred the

kanamycin-resistant gene to the bacterial cells which allowed the colonies to survive the

kanamycin medium.

Conclusion

The ΦOT8 is an effective bacteriophage for generalized transduction in the two Serratia

marcescens strains SM6 and SR41-8000 strains. This can be used by molecular biologists as a

tool to understand deeper the identity and the pathogenic aspect of the S. marcescens.

References

Shirshikova, T.V., Morozova, O.V., Kamaletdinova, L.K., Sharipova, M.R., & Bogomolnaya

L.M. (2016). Generalized Bacteriophage Transduction in Serratia marcescens.

BioNanoScience, 6, 487-489. doi: 10.1007/s12668-016-0268-z