OLEH SUDRAJAT FMIPA UNMUL 2009

Struktur Sel Bakteri

Genom: Kandungan genetik total pada set haploid kromosom pada eukariot Kromosom tunggal pada bakteri atau pada DNA atau RNA virus Material genetik pada organisme

Genom bakteri terdiri dari kromosom sirkular

Terkondensasi dengan cara supercoiling dan looping membentuk badan nukleoid yang tersusun secara rapat Kromosom bereplikasi di dalam sel dan sel membelah secara biner

Plasmids
Extra chromosomal circular DNAs Ditemukan pada bakteri Ukuran bervariasi dari ~ 3,000 bp sampai 100,000 bp. Bereplikasi secara otonomi (origin of replication) Dapat mengandung gen resisten Dapat ditransfer dari satu bakteri ke bakteri lainnya Dapat ditransfer kepada kingdom yang berbeda Multicopy plasmids (~ up to 400 plasmids/per cell) Low copy plasmids (1 2 copies per cell) Digunakan sebagai vektor untuk membawa gen yang diinginkan

CYTOPLASMIC STRUCTURES
Nucleoid area of concentrated DNA no nuclear membrane The DNA is single circular double stranded without proteins

 Ribosom sitoplasmatik, bukan organela

STRUKTUR SITOPLASMATIK
attached to

Plasmid Extrachromosomal loops of DNA

some code for drug resistance toxins

Genotypic Characteristics for Identifying Prokaryotes

the use of genotypic testing has increased with the

availability of technology genotypic testing is particularly useful in the case of organisms that are difficult to identify several techniques include
gene probes PCR sequencing rRNA

Genotypic Characteristics for Identifying Prokaryotes

gene probes single stranded DNA that has been labeled with a identifiable tag, such as a fluorescent dye are complementary to target nucleotide sequences

unique in DNA of pathogen

Microbe gene probed

Genotypic Characteristics for Identifying Prokaryotes

If there is a suspicion, based on symptoms or other environmental parameters that indicates that the organism to be identified may be organism A, a single strand of organism As DNA is introduced with a tag attached (such as fluorescent dye). If the introduced DNA binds to the unknown organism, then it is identified as organism A. If it does not bind to the unknown organism, then the unknown is not organism A.

Genotypic Characteristics for Identifying Prokaryotes

PCR: polymerase chain reaction used to detect small amounts of DNA present in a sample (blood, food, soil) the PCR chain reaction is used to amplify the amount of DNA present

Genotypic Characteristics for Identifying Prokaryotes

sequencing ribosomal RNA
of particular use for identifying prokaryotes impossible to

grow in a culture focus is place on the 16S molecules of the RNA because of its size

approximately 1500 nucleotides

once the 16S molecule is sequenced, it can then be

compared to the sequences of known organisms

Machine used to pick colonies containing wanted DNA

Difficulties in Classifying Prokaryotes

historically prokaryotes have been grouped

according to phenotypic attributes problems with this approach include

mutation resulting in phenotypic changes just because they look alike, does not mean that they are even closely related according the prokaryotics

new molecular approaches are providing better

insight to the relatedness of microorganisms the more similar the nucleotide sequence, the more closely related
DNA extraction

Genotypic Characteristics used in Classifying Prokaryotes

comparison of nucleotide sequences
differences in DNA sequence can assist in determination

of divergence of evolutionary path for organisms DNA hybridization single strands of DNA anneal 16S ribonucleic acid comparing sequence of ribosomal RNA relatedness to other organisms can be determined using numerical taxonomy determined by the percentage of characteristics two organisms have in common The more you have in common phenotypically with another organism the closer related you are to that organism.

Genotypic Characteristics for Identifying Prokaryotes

PCR: polymerase chain reaction used to detect small amounts of DNA present in a sample (blood, food, soil) the PCR chain reaction is used to amplify the amount of DNA present