Клонирование и анализ генов легких цепей иммуноглобулинов стерляди

Клонирование и анализ генов легких цепей иммуноглобулинов стерляди

          V-çîíä âûÿâëÿë áîëåå 20 ãèáðèäèçóþùèõñÿ ôðàãìåíòîâ ÄÍÊ, â òî âðåìÿ êàê Ñ-çîíä ãèáðèäèçîâàëñÿ òîëüêî ñ 3-4 ôðàãìåíòàìè (ðèñ. 13).

           5' íåòðàíñëèðóåìàÿ îáëàñòü      |> ëèäåðíûé ïåïòèä

           1            15              30              45              60              75              90

    B1     . .TTCATGGCAACA GAATCCACAACAACA ATGACTTTTATCAGC ATCTTCATCTGGGCA CTTGTCATCTGCACT CAGGAATCCAGTGGA

    E4     CCC----A------- --------------- --------------- --------------- --------------- ---------------   

    C2     . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .     

    C2(T3) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .     

    PCR    . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .    

          |> V ñåãìåíò   FR 1                                                       |> CDR 1

                       105             120             135             150             165             180

    B1     CAGTATACTGTGACT CAGACTCCAGCAGAG AAATCTGTTCTCCCA GGAGACACAGTCGCT CTCAACTGTAAAGTC AACAGCGCAGTCCTT

    E4     --------------- --------G----T- --------------- ----------. . . . . . . . . . . . . . . . . . .   

    C2     . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .     

    C2(T3) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .     

    PCR    . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

                              |> FR 2                                      |> CDR 2                 |> FR 3

                       195             210             225             240             255             270

    B1V    GGTAATACGTACCTG CACTGGTACCAACAG AAACCTGGAGAAGCT CCCAAACTCCTGATC TATAGGGCAAGTACC CTTGAGTCTGGGATC  

    E4     . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .   

    C2     . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  ---- ---C-----------

    C2(T3) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .     

    PCR    . . . . . . . . . . . . . . . . . .------------ -----------A--- ---TAT----T--AG ---C-------N---

                       285             300             315             330             345             360

    B1     CCAACTCGTTTCAGT GGCAGTGGATCTGGG ACTGACTTCACTCTG ACCATCAGTGGAGTC CAGGCTGAAGATGAA GGAGATTACTACTGT

    E4     . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .   

    C2     --------C----C- --------------- --------------- --------------- --------------- --------T------

    C2(T3) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .   

    PCR    --------------- ------------TT- --------------- --------------- -------------C- ---------------

          |> CDR 3                            |> J ñåãìåíò                                |> Ñ ñåãìåíò

                       375             390             405             420             435             450

    B1     GTCAGTGTACACTAC TCCAGCAGTAGATAT GTGTTGACTTTCGGG CCCGGGACCAAACTA ATTGTGAAATCTGGA AGCCCCACTGCTCCT

    E4     . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  

    C2     CAG---TC------- C----------C-G- -NCC-C--------A --A--------G--G G-------------- -A-------------

    C2(T3) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .   

    PCR    CAG---CAC---ACT C---ATG . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

                       465             480             495             510             525             540

    B1     TCCTCCGTCTCCCTG CTTCCTCCCTCCAAG CTGGAGCTCGACTCA AAGGGTAAAGCCACC CTGGTCTGCCTGGTG AATAATTTCTACCCT

    E4     . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

    C2     --------------- --------------- ------------A-- -----C--------- TG------------- --------------C

    C2(T3) . . . . . . . . . . . . . . . . . . . . . . . . . . -C--------- T-------------- --------------C

    PCR    . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

                       555             570             585             600             615             630

    B1     GATGTCGTGGATATC AAGTGGACGGTGGAC GGGGTGGCCCAGTCG TCTGGTGTGCTGACC AGCACAATGAAGCAG AAGGATGGGAAATAC

    E4     . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

    C2     --------------- --------------T ----C---------- --------------- --------------- -----C---------

    C2(T3) --------------- --------------T ----C---------- --------------- --------------- -----C---------

    PCR    . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

                       645             660             675             690             705             720

    B1     AGTGCAAGCAGCAGC CTGACCCTCACCAAG GCCGTGTGGAACAGC AAGGAGACATACACC TGCACTGTGAAGCAC GAGGCTGTGAGCACT

    E4     . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

    C2     --------------- --------------- -A------------- --------------- --------------- ---------------

    C2(T3) --------------- --------------- -A------------- --------------- --------------- ---------------

    PCR    . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

                                                                 |> 3' íåòðàíñëèðóåìàÿ îáëàñòü

                       735             750             765             780          792 

    B1     CCCAGGAGCGAGTCC ATCAAGAGGAGCGAG TGCACACTATTAGAT GCCtaaAGG . . . . . . . . .

    E4     . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

    C2     --G----------T- --------------- --------------- ---taaG-CAAGAGA ATCCGTGTGATT

    C2(T3) --G----------T- --------------- --------------- . . . . . . . . . . . . . . 

    PCR    . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

          Ðèñ. 11. Íóêëåîòèäíàÿ ïîñëåäîâàòåëüíîñòü ïîëíîðàçìåðíîé êÄÍÊ êëîíà Â1 è ÷àñòè÷íàÿ ïîñëåäîâàòåëüíîñòü êÄÍÊ êëîíîâ Å4 è Ñ2 è ïðîäóêòà ÏÖÐ. Ïîñëåäîâàòåëüíîñòü êÄÍÊ êëîíà Ñ2 îïðåäåëÿëàñü ñ èñïîëüçîâàíèå ïðàéìåðîâ V1, V2, C1 (îáîçíà÷åíèå Ñ2) è Ò3 (îáîçíà÷åíèå Ñ2(Ò3)). Ïðî÷åðê îçíà÷àåò èäåíòè÷íîñòü â ýòîé ïîçèöèè ñ êÄÍÊ êëîíà Â1, FR - êàðêàñíûå ó÷àñòêè, CDR - ãèïåðâàðèàáåëüíûå ó÷àñòêè V îáëàñòè.

A

AruIgLBV     QYTVTQTPAEKSVLPGDTVALNCKVNSAVLGNTYLHWYQQKPGEAPKLLIY*RASTLESGIPTRFSGSG**SGTDFTLTISGVQAEDEGDYYCVSV

IpuIgLVG     -V-------V--A---E--TI--RT-P-Y*-H*-----------------K*F-NQ-H----A------**--S----------T--A-----Q-Y  71%

OmyIgLV1     -I------EM-AFQT--A-T-R-RF-KPSPPC**VA-------G--Q----*Y-T--Q--T-S------**--S-------------A-----Q-Y  63%

XIaIgLVR     -VVL--S-DYV--S--E--TIT--AS-SS---**-------S-QT------*GT-NRYT-T-E------**----------RME---AA----QQY  58%

Rabbit kV IV  ivm----ss---pv----ti--qasqs-ys-nr-a-f-----qp------*k----a--v-s--k---**---q------d--ca-aat---rva  57%

GciIgLVIII     -M--S-PVL--GL-Q--TIT-TASQS-YS-**-A----RE-QK-S----*A-TNRYTEVSE------**---S-----RN--P--VA----QGT  48%

HulV2        -SAL--*-ASV-GS--QSITIS-TGT-SSYNL**VS----H--K----M--*EG-KRP--VSN-----K**--NTAS--T--L-----A----C-Y  47%

HfrIgLVII    GTVL--*--SM-TSQ-K--KIT-TISGGGTYY**SS--W----S--VFVWRDYD--RG----D--T--RNT-SNVMH---TD--SR-TA-----AW  41%

XlaIgLIII    -VSI--*-VSE--KL-E--RIS-TLSG-SGYH**VN-----A-NR-RY-LRFYSDSNK**HQD-----KDSPNNIGY---K-ALL--DA----ATW  35%

HfrIgLV1     VPVLN---ISDP-SA-E-SE-K-AMQNGGSYY**MS--R-R-----VFVL**YQ--SG-IYRD--KP-RDT-SNSHI---GSLEPG-SAV---AAN  31%

Á

AruIgLCB    GSPTAPSSVSLLPPSKLELDSKGKATLVCLVNNFYPDVVDIKWTVDGVAQSSG*VLTSTMKQK**DGKYSASSSLTLTKAVWNSKETYTCTVKHEAVSTPRSESIKRSECTLLDA

CplIgLCII     -RS*PT--V----SDQITA-NM-------SG-V-GAAE-E-----SVRGN-*-E--RIQ-EA*-NTF-V--Y---SASD---H-L-S-V-K-ETQAN-LQT--S--S-M     44%

HfrIgLCIII    EKSQPTLT-M---PE-VKA--T------ADH----E-GVE-KK--A-I-A-*-Q--NYLRAS*-ST--C--L---SGSD-E-NARFS-ALT-ETL-S-L-K-VS----V     42%

IpuIgLCF     G--VKP-----L--S-Q-S*E-S-S-L--LPAYS-QGALVS-----SEVKD-*----AEER-**TDG-TR--T---S--L-EKG-EFV-K-S-DN-DH-VT**FRK-Q-EV    41%

Hu kappa      TV-AP-VFIF---*D-QLKS-T-SV---L-----REAKVQ-K--NAL--GN*SQE-VTE-DSK-ST--L--T---S--DYEKHKV-A-E-T-QGL-S-VTK-FN-G--      39%

XlaIgLC3    GDVK--*---YF---*V-EIATK---V--SLSD-T-RGATVK-L---KD-TDS*-QS-GLSKQS*-NL-ME--Y-S--ADQ-LRH---S-K-S-Q**GKEIIQTL-----V     38%

Hu lambda1   Q-K-NPT-T-F---*S-ELQAN-------ISD---GA-TVA-KA--SPVKA-*-E-TKPSKQS*NN--A---Y-S--PEQ-K-HRS-S-Q-T-E**GSTVEKTVAPT--S     37%

HfrIgLCI     SEDRKP-VL-----*S-EIDS-W---S---SR-K-GF-RVL-R--DKETD--*-T-G-VSTDS*-QS--L--YLRVPATA--KGSS---S-D-GSL-S-LLKT-SSTA-SD    37%

XlaIgLCS      ND-KPA-FIFK--*D-QVKE-NP-A---I---F-RDLTVT-K--SQDV--SD-K--DFM-ES*-ST--Q--M-T---DK-DKADKFE-L-K-K**TAQLTQSFSK-Q-S     34%

IpuIgLCG       LTQP--TV----SV--Q*QE-V-----AYKGF-SDWRLS-K---SSW---*ESR-SAVLQA*--L--W--T-S-HPEQ-RN-*VV--EASKDN*QP-VVSTVNTEQ-      33%

          Ðèñ. 12. Ñðàâíåíèå àìèíîêèñëîòíûõ ïîñëåäîâàòåëüíîñòåé V (À) è Ñ (Á) îáëàñòåé L-öåïè Èà ñòåðëÿäè ñ ïîñëåäîâàòåëüíîñòÿìè äðóãèõ âèäîâ ïîçâîíî÷íûõ. Äåôèñ îáîçíà÷àåò èäåíòè÷íîñòü â äàííîé ïîçèöèè. Çâåçäî÷êà îáîçíà÷àåò äåëåöèþ àìèíîêèñëîòíûõ îñòàòêîâ. Ñëåâà óêàçàíû ïðîöåíòû ñõîäñòâà. Âèäîâûå îáîçíà÷åíèÿ: Aru - ñòåðëÿäü, Ipu - ïåùåðíûé ñîìèê, Omy - ðàäóæíàÿ ôîðåëü, Xla - øïîðöåâàÿ ëàãóøêà, Gci - àêóëà-íÿíüêà, Hu - ÷åëîâåê, Cpl - ïåñ÷àíàÿ àêóëà, Hfr - ðàçíîçóáàÿ àêóëà.

          Ðèñ. 13. Ñàóçåðí áëîò ãèáðèäèçàöèÿ. Ãåíîìíàÿ ÄÍÊ áûëà âûäåëåíà èç ïå÷åíè è îáðàáîòàíà ýíäîíóêëåàçàìè ðåñòðèêöèè Pst I Pvu II. Ãèáðèäèçàöèþ ïðîâîäèëè â ìÿãêèõ óñëîâèÿõ ñ èñïîëüçîâàíèåì çîíäîâ, ãîìîëîãè÷íûõ VL (à) è CL (á) ãåííûì ñåãìåíòàì L-öåïåé Èà ñòåðëÿäè. Ñ V-çîíäîì ãèáðèäèçîâàëîñü áîëåå 20 ôðàãìåíòîâ ÄÍÊ, â òî âðåìÿ êàê ñ Ñ-çîíäîì ãèáðèäèçîâàëîñü òîëüêî 3-4 ôðàãìåíòà.

ÎÁÑÓÆÄÅÍÈÅ ÐÅÇÓËÜÒÀÒÎÂ

           ðåçóëüòàòå ïðîâåäåííîé ðàáîòû èäåíòèôèöèðîâàí ëîêóñ ãåíîâ, êîäèðóþùèõ L-öåïè Èà ñòåðëÿäè, ïðåäñòàâèòåëÿ ïîäêëàññà êîñòíî-õðÿùåâûõ ðûá. Ñðàâíèòåëüíûé àíàëèç ñòðóêòóðû V ãåíîâ ýòîãî ëîêóñà óêàçûâàåò, ÷òî îí îòíîñèòñÿ ê êàïïà òèïó. Ðåçóëüòàòû ñðàâíèòåëüíîãî àíàëèçà Ñ îáëàñòåé ìåíåå èíôîðìàòèâíû, ÷òî ñàìî ïî ñåáå íå óäèâèòåëüíî, òàê êàê Ñ-ãåíû L-öåïåé Èà çíà÷èòåëüíî ìåíåå êîíñåðâàòèâíû ÷åì V-ãåíû. Òåì íå ìåíåå, òîò ôàêò, ÷òî íà íóêëåîòèäíîì óðîâíå Ñ-ãåí ñòåðëÿäè èìååò íàèáîëüøåå ñõîäñòâî ñ êàïïà-ïîäîáíûìè ãåíàìè III êëàññà õðÿùåâûõ ðûá, òàêæå ñâèäåòåëüñòâóåò î òîì, ÷òî îáíàðóæåííûé ëîêóñ îòíîñèòñÿ ê êàïïà òèïó.

          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 Èà 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 È. È. Èõòèîëîãèÿ. Ìîñêâà-Ëåíèíãðàä: Ïèùåïðîìèçäàò, 1941.

          Aguilera R. J., Akira S., Okazaki K.,Sakano H. A pre-B cell nuclear protein wich specifically interacts with the immunoglobulin V-J recombination sequeces // Cell. 1987. V. 51. P. 909-917.

          Anderson M. K., Shamblott M. J., Litman R. T., Litman G. W. Generation of immunoglobulin light chain gene diversity in Raja erinacea is not associated with somatic rearrangement, an exception to a central paradigm of B cell immunity // Journal of Experimental Medecine. 1995. V. 182. P. 109-119.

          Bengten E. The immunoglobulin genes in Atlantic cod (Gadus morhuna) and rainbow trout (Oncorhynchus mykiss) // Dissertations from the Faculty of Science and Technology. 1994. V. 19.

          Bengten E., Leanderson T., Pilstrom L. Immunoglobulin heavy chain cDNA from Atlantic cod (Gadus morhuna L.): nucleotide sequences of secretory and membrane form show an unusual splicing pattern // European Journal of Immunology. 1991. V. 21. P. 3027-3033.

          Blomberg B., Tonegawa S. DNA sequences of the joining regions of mouse l light chain immunoglobulin genes // Proc. Natl. Acad. Sci. USA. 1982. V. 79. P. 530.

          Boyle J. S., Lew A. M. An inexpensive alternative to glassmilk for DNA purification // Trends in Genetics. 1995. V. 11. No. 1. p. 8.

          Chomczynski P. and Sacchi N. Single-step method of RNA isolation by acid guanidinum thiocyanate-phenol-chloroform extraction // Analytical biochemistry. 1987. V. 162. P. 156-159.

          Daggfeldt A., Bengten E., Pilstrom L. A claster type organization of the loci of the immunoglobulin light chain in Atlantic cod (Gadus morhuna L.) and rainbow trout (Oncorhynchus mykiss Walbaum) indicated by nucleotide sequences of cDNAs and hybridization analysis // Immunogenetics. 1993. V.38. P. 199-209.

          Dieffenbach C. W. and Dveksler G. S. (edit). PCR primer. A laboratory manual. New York: Cold Spring Harbor Laboratory Press, 1995. 714 p.

          Durdik J., Moore M. W., Selsing E. Novel k light-chain gene rearrangements in mouse l light-chain-producing B limphocytes // Nature. 1984. V. 307. P. 749-752.

          Flajnik M. F. The immune system of ectothermic vertebrates // Veterinary Immunology and immunopathology. 1996. V. 54. P. 145-150.

          Ghaffari S. H. and Lobb C. J. Structure and genomic organization of immunoglobulin light chain in the channel catfish // The Journal of Immunology. 1993. V. 151. P. 6900-6912.

          Gilbert W. Evolution of antibodies: the road not taken // Nature. 1990. V. 320. P. 485.

          Greenberg A. S., Steiner L.,Kasahara M., Flajnik M. F. Isolation of a immunoglobulin light chain cDNA clone encoding a protein resembling mammalian k light chains: Implications for the evolution of light chains // Proc. Natl. Acad. Sci. USA. 1993. V. 90. P. 10603-10607.

          Haire R. N., Ota T., Rast J. P., Litman R. T., Chan F. Y., Zon L. I., Litman G. W. A third Ig light chain gene isotype in Xenopus laevis consists of six distinct VL families and is related to mammalian l genes // The Journal of Immunology. 1996. V. 157. P. 1544-1550.

          Hesse J. E., Lieber M. R., Mizuuchi K., Gellert M. V(D)J recombination: the functional difinition of the joining signals // Genes Dev. 1989. V. 3. P. 1053-1061.

          Hieter P. A., Hollis G. F., Korsmeyer S. J., Waldmann T. A., Leder P. Clastered arrangement of immunoglobulin l constant region genes in man // Nature. 1981. V. 294. P. 536.

          Hieter P. A., Maizel J. V., Leder J., Leder P. Evolution of human immunoglobulin k J region genes // J. Biol. Chem. 1982. V. 257. P. 1516.

          Hohman V. S., Schluter S.F., Marchalonis J. J. Complete sequence of a cDNA clone specifying sandbar shark immunoglobulin light chain: Gene organization and implications for the evolution of light chains // Proc. Natl. Acad. Sci. USA. 1992. V. 89. P. 276-280.

          Hohman V. S., Schuchman D. B., Schluter S.F., Marchalonis J. J. Genomic clone for sandbar shark l light chain: Generation diversity in the absence of rearrangement // Proc. Natl. Acad. Sci. USA. 1993. V. 90. P. 9882-9886.

          Hsu E., Lefkovits I., Flajnik M., Pasquier L. D. Light chain heterogeneity in the amphibian Xenopus // Molecular Immunology. 1991. V. 28. P. 985-994.

immunoglobulin genes and the antibody repertoire // Molecular Biology Evolution. 1993. V. 10(1). P. 60-72.

          Litman G. W., Rast J. p., Shamblott M. J., Haire R. N., Michele H., Roess W., Litman R. T, Hinds-Frey K. R., Zilch A., Amemiya C. T. Phylogenetic diversification of    Lobb C. J., Olson M. O. J., Clem W. L. Immunoglobulin light chain classes in a teleost fish // The Journal of Immunology. 1984. V. 132. P. 1917-1923.

          Lundqvist M., Bengten E., Stromberg S., Pilstrom L. Ig light chain gene in the siberiah sturgeon (Acipenser baeri) // The Journal of Immunology. 1996. V. 157. P. 2031-2038.

          Maisey J. G. Chondrichthyan phylogeny: a look at the evidence // J. Ven. Paleont. 1984. V. 4. P. 359-371.

          Rast J. P., Anderson M. k., Ota T. Litman R. T., Margittai M., Shamblott M. J., Litman G. W. Immunoglobulin light chain class multiplisity and alternative organizational form in early vertebrate philogeny // Immunogenetics. 1994. V. 40. P. 83-89.

          Reynaund C. A., Anquenz V., Grimal H., Weill J. C. A hyperconversion mechanism generates the chicken light chain preimmune repertoire // Cell. 1987. V. 48. P. 379-388.

          Roitt I., Brodstoff J., Male D. Immunology. London: Mosby-Year Book Europe, 1993.

          Romer A. S. Vertebrate Paleontology. Chicago: University Chicago Press, 1966.

          Sakano H., Huppi K., Heinrich G., Tonegawa S. Sequences at the somatic recombination sites of immunoglobulin light-chain genes // Nature. 1979. V. 280. P. 288.

          Sakano H., Maki R., Kurosawa Y., Roeder W., Tonegawa S. Two types of somatic recombination are necessary for the generation of complete immunoglobulin heavy-chain genes // Nature. 1980. V. 286. P. 676.

          Sanger F., Niklen, S., Coulson A.R. DNA sequencing with chain-terminating inhibitors // Proc. Natl. Acad. Sci. U S A. 1977. V. 74. P. 5463-5467

          Schatz G. D., Oettinger A. M., Schlissel M. S. V(D)J recombination: molecular biology and regulation // Annual Review of Immunology. 1992. V. 10. P. 359-383.

          Schwager J., Burckert N., Schwager M., Wilson M. Evolution of immunoglobulin light chain genes: analysis of Xenopus IgL isotypes and their contribution to antibody diversity // The EMBO Journal. 1991. V. 10. P. 505-511.

          Seidman J. G., Max E. E., Leder P. A. A k immunoglobulin gene is formed by site - specific recombination without further somatic mutation // Nature. 1979. V. 280. P. 370.

          Shamblott M. J. and Litman G. W. Genomic organization and sequences of immunoglobulin light chain genes in a primitive vertebrate suggest coevolution of immunoglobulin gene organization // The EMBO Journal. 1989. V. 8. P. 3733-3739.

          Shankey T.V. and Clem L.W. Phylogeny of immunoglobulin structure and function. IX. Intramolecular heterogeneity

of shark 19S IgM antibodies to the dinitrophenyl hapten //

J. Immunol. 1980. V. 125. P. 2690-2698.

          Stavnezer J., Kekish O., Batter D., Grenier J., Balazs I., Henderson E., Zegers B. J. Aberrant recombination events in B cell lines derived from k-deficient human // Nucl. Acids Res. 1985. V. 13. P. 3495-3514.

          Stewart S. E., Pasquier L. D., Steiner L. A. Diversity of expressed V and J regions of immunoglobulin light chains in Xenopus laevis // European Journal of Immunology. 1993. V. 23. P. 1980-1986.

          Strob U. Transgenic mise with immunoglobulin genes // Annual Review of Immunology. 1987. V. 5. P. 151-174.

          Thompson G. B. and Neiman P. E. Somatic diversification of the chicken immunoglobulin light chain gene is limited to the rearranged variable gene segment //Cell. 1987. V. 48. P. 369-378.

          Tonegawa S. Somatic generation of antibody diversity // Nature. 1983. V. 302. P. 575-581.

          Udey J. A., Blomberg B. Human l light chain locus: organization and DNA sequences of three genomic J regions // Immunogenetics. 1987. V. 25. P. 63.

          Wai Lin Tung, King-C. Chow. A modified medium for efficient electrotransformation of E.coli // Trends in Genetics. 1995. V. 11. No. 4. Ð. 128-129.

          Wilson M., Hsu E., Marcuz A., Courent M., Pasquier L. D., Steinberg C. What limits affinity maturation of antibodies in Xenopus: the rate of somatic mutation or the abilityto select mutants? // The EMBO Journal. 1992. V. 11. P. 4337.

          Zezza D. J., Mikoryak C. A., Schwager J., Steiner L. A. Sequence of C region of L chains from Xenopus laevis Ig // The Journal of Immunology. 1991. V. 146. P. 4041-4047.

          Zezza D. J., Stewart S. E., Steiner L. A. Genes encoding Xenopus laevis Ig L chains. Implications for the evolution of k and l chains // The Journal of Immunology. 1992. V. 149. P. 3968-3977.

Страницы: 1, 2, 3, 4, 5, 6, 7