Клонирование и анализ генов легких цепей иммуноглобулинов стерляди 
êÄÍÊ
êëîíîâ Â1, Ñ2, Å4,
à òàêæå
ôðàãìåíò,
ïîëó÷åííûé
ñ ïîìîùüþ ÏÖÐ,
ñîäåðæàò
áëèçêîðîäñòâåííûå
V-ãåíû.
Ðàçëè÷èÿ ìåæäó
íèìè
ñêîíöåíòðèðîâàíû
â îñíîâíîì â
îáëàñòè 3-ãî
ãèïåðâàðèàáåëüíîãî
ðàéîíà. Ïîñëåäîâàòåëüíîñòè
äâóõ
îáíàðóæåííûõ
J-ñåãìåíòîâ
îòëè÷àþòñÿ
ïî 10
íóêëåîòèäàì,
ò.å., îíè
ïðåäñòàâëÿþò
ñîáîé
ðàçëè÷íûå
ãåíîìíûå
ñåãìåíòû.
Íåçíà÷èòåëüíûå
ðàçëè÷èÿ îáíàðóæåíû
òàêæå ìåæäó
òðåìÿ
ïîñëåäîâàòåëüíîñòÿìè
Ñ-ãåíîâ.
Ñ-ñåãìåíò
êëîíà Â1 îòëè÷àåòñÿ
îò ïîëíîãî
Ñ-ñåãìåíòà
êëîíà Ñ2 ïî 9
ïîçèöèÿì, èç
êîòîðûõ 5
âåäóò ê
çàìåíå àìèíîêèñëîòû.
Ïî-âèäèìîìó,
ýòè
ïîñëåäîâàòåëüñòè
òàêæå
ïðåäñòàâëÿþò
ñîáîé
ðàçíûå
çàðîäûøåâûå
ãåíû.
Àíàëèç
ñòðóêòóðíîé
îðãàíèçàöèè
ëîêóñà ñ
ïîìîùüþ
Ñàóçåðí
áëîò-ãèáðèäèçàöèè
óêàçûâàåò,
÷òî ó
ñòåðëÿäè
èìåþòñÿ
ìíîæåñòâåííûå
VL ãåíû.
Òî÷íàÿ
îöåíêà èõ
êîëè÷åñòâà
íåâîçìîæíà,
íî íàëè÷èå
áîëåå 20 ïîëîñ
ãèáðèäèçàöèè
ñ ðàçíîé
èíòåíñèâíîñòüþ
ñèãíàëà, ïî àíàëîãèè
ñ
ìíîãî÷èñëåííûìè
ëèòåðàòóðíûìè
äàííûìè,
ïîçâîëÿåò
ãîâîðèòü î
íåñêîëüêèõ
äåñÿòêàõ
êîïèé.
Ïðè
èñïîëüçîâàíèè
Ñ-çîíäà
êîëè÷åñòâî
ïîëîñ â
ìÿãêèõ
óñëîâèÿõ
ãèáðèäèçàöèè
íå ïðåâûøàåò
4-õ. Ó÷èòûâàÿ
âîçìîæíîñòü
àëëåëèçìà
ìîæíî ãîâîðèòü
î íàëè÷èè â
ýòîì ëîêóñå
2-4õ ãåíîâ.
ßðêî
âûðàæåííîå
êîëè÷åñòâåííîå
ïðåâîñõîäñòâî
V ãåíîâ íàä
Ñ-ãåíàìè
ÿâëÿåòñÿ
õàðàêòåðíûì
ïðèçíàêîì
ñåãìåíòàðíîé
ôîðìû îðãàíèçàöèè
ëîêóñà. Ó
õðÿùåâûõ è
êîñòèñòûõ
ðûá ñ
êëàñòåðíîé
îðãàíèçàöèåé
ãåíîâ L-öåïåé
êîëè÷åñòâî V
è Ñ ãåíîâ
î÷åíü âåëèêî
è
ïðèáëèçèòåëüíî
îäèíàêîâî,
÷òî îòðàæàåòñÿ
â ñõîäíîé
êàðòèíå
áëîò-ãèáðèäèçàöèè.
Ïîëó÷åííûå
íàìè
ðåçóëüòàòû
ÿâëÿþòñÿ ñåðüåçíûì
îñíîâàíèåì
äëÿ òîãî,
÷òîáû óòâåðæäàòü,
÷òî ó
ñòåðëÿäè
îðãàíèçàöèÿ
ãåíîâ L-öåïåé
êàïïà-ïîäîáíîãî
òèïà èìååò ñåãìåíòàðíûé
õàðàêòåð,
ïîäîáíûé
îðãàíèçàöèè
êàïïà
ëîêóñà
ìëåêîïèòàþùèõ.
Ýòè
äàííûå
óêàçûâàþò
íà òî, ÷òî
ïåðåõîä îò
êëàñòåðíîé
ê
ñåãìåíòàðíîé
îðãàíèçàöèè
ïðîèçîøåë â
ôèëîãåíåçå
ïîçâîíî÷íûõ
î÷åíü ðàíî,
âîçìîæíî
óæå ó
äðåâíèõ
ïåðâè÷íî-êîñòíûõ.
ÂÛÂÎÄÛ
1.
Ñêîíñòðóèðîâàíà
áèáëèîòåêà
êÄÍÊ ëåéêîöèòîâ
ñòåðëÿäè
ïðåäñòàâèòåëüíîñòüþ
6õ106
íåçàâèñèìûõ
êëîíîâ. ÏÖÐ
ÄÍÊ
áèáëèîòåêè ñ
ïîìîùüþ
âûðîæäåííîãî
ïðàéìåðà,
ñîîòâåñòâóþùåãî
J ñåãìåíòó
ãåíîâ ÈÃ,
ïîçâîëèë âûÿâèòü
ôðàãìåíò ÄÍÊ
äëèíîé 370 ïí,
ãîìîëîãè÷íûé
V ãåíàì ÈÃ
ïîçâîíî÷íûõ.
2.
Èç
áèáëèîòåêè
êÄÍÊ ñ
èñïîëüçîâàíèåì
ïðîäóêòà ÏÖÐ
â êà÷åñòâå
çîíäà
âûäåëåíî
ïÿòü êëîíîâ,
ñîäåðæàùèõ
âñòàâêè
êÄÍÊ ãåíîâ
L-öåïåé ÈÃ.
Ñðàâíèòåëüíûé
àíàëèç
ïåðâè÷íîé ñòðóêòóðû
êÄÍÊ êëîíîâ
ïîêàçàë, ÷òî
îíè ÿâëÿþòñÿ
÷ëåíàìè
îäíîãî
ñåìåéñòâà
ãåíîâ è
èìåþò
íàèáîëüøåå
ñõîäñòâî ñ
ãåíàìè êàïïà
òèïà.
3.
Ñîãëàñíî
ðåçóëüòàì
ãåíîìíîãî
áëîò-àíàëèçà
èäåíòèôèöèðîâàííîå
ñåìåéñòâî
ñîäåðæèò
íåñêîëüêî
äåñÿòêîâ V
ãåíîâ è
òîëüêî 2-4 Ñ
ãåíà, ÷òî
óêàçûâàåò
íà
ñåãìåíòàðíóþ
ôîðìó åãî
îðãàíèçàöèè.
4.
Ïîëó÷åííûå
äàííûå
óêàçûâàþò
÷òî ïåðåõîä
îò
êëàñòåðíîãî
òèïà
îðãàíèçàöèè
ãåíîâ ÈÃ,
õàðàêòåðíîãî
äëÿ õðÿùåâûõ
ðûá, ê ñåãìåíòàðíîìó,
ïðèñóòñòâóþùåìó
ó âñåõ ìëåêîïèòàþùèõ,
ïðîèçîøåë â
ýâîëþöèè â
ïåðèîä ïîÿâëåíèÿ
ïåðâè÷íî-êîñòíûõ
ðûá.
ÑÏÈÑÎÊ
ËÈÒÅÐÀÒÓÐÛ
Ìàçèí
À. Â.,
Êóçíåäåëîâ
Ê. Ä. è äð.
Ìåòîäû ìîëåêóëÿðíîé
ãåíåòèêè è
ãåííîé
èíæåíåðèè.
Íîâîñèáèðñê,
1988. 333 ñ.
Ìàíèàòèñ
Ò., Ôðè÷ Ý.,
Ñýìáðóê Ä.
Ìîëåêóëÿðíîå
êëîíèðîâàíèå.
Ì.: Ìèð, 1984. 480 ñ.
Ïîë
Ó. (ðåä.)
Èììóíîëîãèÿ.
Ì.: Ìèð, 1987. Ò. 1. 476 ñ.
Ðîìåð
À. Ñ. è Ïàðñîíñ
Ò. Ñ.
Àíàòîìèÿ
ïîçâîíî÷íûõ.
Ì.: Ìèð, 1992. 358 ñ.
Ôðèìåëü
Ã. (ðåä.)
Èììóíîëîãè÷åñêèå
ìåòîäû. Ì.:
Ìåäèöèíà, 1987. 472
ñ.
Þäêèí
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