Antisense molecular beacon strategy for in situ visualization of snRNA and fibrillarin protein interaction in Giardia lamblia.
Use of confocal microscopy has provided many recent developments in the study of functional aspects, especially localization and distribution of proteins, DNA and RNA within the cells. In the present investigation, we have applied for the first time, antisense molecular beacon based Fluorescence Resonance Energy Transfer (FRET) and Flow Cytometric Energy Transfer (FCET) techniques to demonstrate binding and co-localization of fibrillarin protein with small nuclear RNA (snRNA) to form ribonucleoprotein particle (RNPP) complex in Giardia lamblia. It has been observed by FRET and FCET that energy transfer occurs from fluorescence tagged fibrillarin to snRNA antisense molecular beacon confirming the clear physical interaction between them during RNPP complex formation. This is the first demonstration of in situ detection of RNA-protein complex formation by antisense molecular beacon based FRET and FCET in Giardia lamblia.
Ganguly
Sandipan
S
National Institute of Cholera and Enteric Diseases, Indian Council of Medical Research, Beliaghata, Kolkata, India. sandipanganguly@hotmail.com
Ghosh
Srikanta
S
Chattopadhyay
Dhrubajyoti
D
Das
Pradeep
P
D000818:Animals; D001483:Base Sequence; D002467:Cell Nucleus / Q000378:metabolism; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry*; D004251:DNA Transposable Elements; D031541:Fluorescence Resonance Energy Transfer / Q000295:instrumentation / Q000379:methods*; D005821:Genetic Techniques; D016829:Giardia lamblia / Q000378:metabolism*; D046911:Macromolecular Substances; D018613:Microscopy, Confocal / Q000379:methods; D008969:Molecular Sequence Data; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D012342:RNA, Small Nuclear / Q000737:chemistry*; D012261:Ribonucleoproteins / Q000737:chemistry
D002868:Chromosomal Proteins, Non-Histone; D004251:DNA Transposable Elements; D046911:Macromolecular Substances; D012333:RNA, Messenger; D012342:RNA, Small Nuclear; D012261:Ribonucleoproteins; C047458:fibrillarin