Formation and Investigations of Bioselective Element of Surface Plasmon Resonance Sensor for Recognition of Specific Oligonucleotide Sequences

Abstract

Àíîòàö³ÿ. Á³îñåëåêòèâíèé åëåìåíò ñåíñîðà ïîâåðõíåâîãî ïëàçìîííîãî ðåçîíàíñó (ÏÏÐ) äëÿ ðîçï³çíàâàííÿ îë³ãîíóêëåîòèäíèõ ïîñë³äîâíîñòåé, ïîâ'ÿçàíèõ ç ã³áðèäíèì ãåíîì bcrabl, ñòâîðåíî øëÿõîì ³ììîá³ë³çàö³¿ íà ñåíñîðí³é ïîâåðõí³ ò³îëüîâàíèõ îäíîëàíöþãîâèõ îë³-ãîíóêëåîòèä³â òà 6-ìåðêàïòî-1-ãåêñàíîëó. Îë³ãîíóêëåîòèä ç 24 îñíîâ, ÿêèé ðåïðåçåíòóº ä³-ëÿíêó ñòèêóâàííÿ ã³áðèäíîãî ãåíà ³ ñêëàäàºòüñÿ ç îäíàêîâèõ çà äîâaeèíîþ ôðàãìåíò³â îáîõ âèõ³äíèõ ãåí³â, áóâ âèêîðèñòàíèé ÿê ïðîáà äëÿ ³ììîá³ë³çàö³¿ íà ñåíñîðí³é ïîâåðõí³. ³äá³ð ïîñë³äîâíîñò³ öüîãî îë³ãîíóêëåîòèäó áàçóâàâñÿ íà GC ñêëàä³ òà çäàòíîñò³ äî âíóòð³øíüî-òà ì³aeìîëåêóëÿðíèõ âçàºìîä³é, òåîðåòè÷íî ïåðåäáà÷åíî¿ çà äîïîìîãîþ âåá-ñåðâåðà DINAMelt. Ôîðìóâàííÿ á³îñåëåêòèâíîãî åëåìåíòó ³ íàñòóïí³ åòàïè ã³áðèäèçàö³¿ ç êîìïëåìåíòàðíèìè îë³ãîíóêëåîòèäàìè êîíòðîëþâàëè â ðåaeèì³ ðåàëüíîãî ÷àñó, êîðèñòóþ÷èñü åêñïåðèìåíòàëü-íèì ìàêåòîì ñïåêòðîìåòðà "Ïëàçìîí ÏÏÐ-4ì", ðîçðîáëåíîãî â ²íñòèòóò³ ô³çèêè íàï³âïðî-â³äíèê³â ÍÀÍ Óêðà¿íè. Áóëî ïîêàçàíî, ùî ò³îëüîâàí³ îäíîëàíöþãîâ³ îë³ãîíóêëåîòèäè modPh óñï³øíî ³ììîá³ë³çóþòüñÿ íà çîëîò³é ïîâåðõí³ âèì³ðþâàëüíî¿ êîì³ðêè ñïåêòðîìåòðà ÏÏÐ ó 0,5 M KH 2 PO 4 (ðÍ 3,8). ̳ae ³ììîá³ë³çîâàíèìè mod-Ph òà êîìïëåìåíòàðíèìè îë³ãîíóêëåî-òèäàìè P1 â³äáóâàëàñÿ ñïåöèô³÷íà ã³áðèäèçàö³ÿ, â òîé ÷àñ ÿê ââåäåííÿ íåêîìïëåìåíòàðíèõ îë³ãîíóêëåîòèä³â ñåíñîðíîãî â³äãóêó íå âèêëèêàëè. Åêñïåðèìåíòàëüí³ ñåíñîðí³ â³äãóêè, îò-ðèìàí³ ïðè âçàºìîä³ÿõ äîñë³äaeóâàíèõ îë³ãîíóêëåîòèä³â íà ñåíñîðí³é ïîâåðõí³ äîñèòü äîáðå óçãîäaeóþòüñÿ ç òåîðåòè÷íèìè ðîçðàõóíêàìè òåðìîäèíàì³÷íèõ ïàðàìåòð³â öèõ âçàºìîä³é.Êëþ÷îâ³ ñëîâà: ïîâåðõíåâèé ïëàçìîííèé ðåçîíàíñ, ã³áðèäèçàö³éíèé ÄÍÊ-ñåíñîð, á³îñåëåêòèâíèé åëåìåíò, ãåí bcr-abl FORMATION AND INVESTIGATIONS OF BIOSELECTIVE ELEMENT OF SURFACE PLASMON RESONANCE SENSOR FOR RECOGNITION OF SPECIFIC OLIGONUCLEOTIDE SEQUENCESA. E. Rachkov, Yu. V. Holodova, M. V. Dybkov, G. D. Telegeev, A. P. Soldatkin Abstract. A bioselective element of the surface plasmon resonance (SPR) sensor for recognition of oligonucleotide sequences related to the hybrid gene bcr-abl was prepared by immobilization of thiol-derivatized single-stranded oligonucleotides and 6-mercapto-1-hexanol on the sensor surface.