A Seminar On

Genetic Recombination in Bacteria

Supervisor Dr. P.K. Rai

Submitted By
Sapana

INTRODUCTION
Genetic recombination is the formation of a new genotype by exchange of genetic material between two different chromosome which have similar gene at corresponding sites.

The prokaryotes have evolved different mechanism for creating recombinants. These mechanisms are collectively called as Horizontal gene transfer (HGT).

In Bacteria, genetic recombination occur from three types of horizontal gene transfer.
1. Conjugation Transfer of genes between cells that are in physical contact with each other. Transduction Transfer of genes from one cell to another is mediated by a bacteriophage. Transformation Transfer of naked DNA or cell free from one cell to another.

2.

3.

Genetic Recombination in bacteria

The three types of gene transfer in bacteria are as follows Conjugation Transformation Transduction Conjugation Lederberg and Tatum (1946) discovered conjugation in E.coli and its detailed studies were made by woolman and Jacob (1956).

1. 2. 3. 1.

Evidence of Bacterial conjugation

1. Lederberg and Tatums mixed two autotrophic strains, incubated the culture for several hours in nutrient medium and then plated it on minimal medium, where only prototrophs could grow. They used one strain required Biotin (Bio-), Phenylalanine (Phe-) and Cystine (Cys-) for growth and the other needed Threonine (Thr-), Leucine (Leu-) and Thiamine (thi-). Bernard Davis (1950) proved that physical contact of cells is necessary for gene transfer he performed a U tube experiment. Plasmids are small double stranded DNA molecules that can exist independently of host chromosomes. They have their own replication origins and autonomously replicate and stably inherited.

2.

Conjugation between a F+ (donor) cell and F(recipient) cell

In 1952 William Hays demonstrated that the gene transfer was polar. That is there were definite donor (F+ or fertile) cell and recipient (For nonfertile) strain and gene transfer was non reciprocal. He also observed that in a F+ Fmating, the chromosomal genes were rarely transferred to progency, but F- strain frequently become F+.

Conjugation between Hfr and F- cell

In this type of conjugation, donor cell transfers chromosomal genes with great efficiency, but does not change the recipient bacteria into F+ cells. Because of high frequency of recombination produced by this mating, it is referred to as Hfr conjugation and the donor is called an Hfr strain. During transfer, the Hfr chromosome begins replicating at the point of insertion of the Ffactor, the Hrf chromosome begins replicating at the point of insertion of the F- factor, the integrated F- factor serves as the point of chromosomal opening and some part of it serves as the origin of transfer.

Conjugation between F (F- prime) male and F- (female) cell When a F cell is conjugate with Frecipient cell the sex-factor is transferred very efficiently together with the added bacterial genes. The recipient cell then becomes a heterozygous or partial diploid, because the some bacterial genes present on the F plasmid are also found on the recipient s chromosome. Jacob and Wollman termed this type of genetic recombination as sex-duction.

Transformation
Bacterial transformation may be referred as a stable genetic change brought about by taking up naked DNA (i.e. DNA without associate cells proteins.) Transformation was first discovered by Griffith in 1928 in streptococcus. Mechanism of transformation In transformation process a naked DNA is transferred from one bacterial cell to another. The DNA is obtained from the donor cell either by natural lysis or by chemical extraction. The DNA is taken in, through the cell membrane of the recipient cell. After DNA entry into recipient cell, one strand of double stranded DNA is degraded by endonuclease enzyme, while the other strand undergoes base pairing with the homologous portion of the recipient DNA with the help of enzyme ligase.

Transduction
Transduction discovered by N. Zinder and J. Lederberg in 1952 in Salmonella typhimurium. In transduction, the fragments of DNA are transferred from one bacterial cell (donor) to other (recipient), with the help of bacteriophage serving as vector. There are two different kinds of transduction generalized and specialized. a) Generalized Transduction : Transduction which results in transfer of any part of bacterial gene from bacterial cell to the other is referred to as Generalized or nonspecialized transduction. It is mediated by some virulent phages and certain temprate phases; E.Coli phage P.I., Salmonella phage P. 22 and Bacillus subtilis phages PBS 1 and SP10 are such phages.

b) Specialized transduction : In contrast to generalized (non-restricted) transduction which results in transfer of any gene from donor to recipient bacterial cell, specialized (restricted) transduction is that which lead to the transfer of only specific (restricted) gene from donor to recipient cell. Specialized transduction is mediated by those terperature bacteria phages (eg Lambda) () phage, Mu () phase and 80 phage) that usually incorporate (integrate) their DNA into the bacterial chromosome.

Conclusion
Biacteria have no sexual reproduction in the sense that the eu-karyotes do. They have no alternation of diploid and haploid generations, no gametes, no meiosis. But the essence of sex is genetic recombination. "Genetic recombination is the formation of a new genotype by reassortment of genes following an exchange of genetic material between two different chromosomes." It may some what surprising that bacteria can undergo recombination because the process of recombination requires two homologous DMA molecules and bacteria have only one chromosome and therefore haploid. The bacterial recombination promote homologous recombination in the laboratory and they have potential to mediate the replace next of regions on the bacterial chromosome with the corresponding regions from other members of same or closely related species.